Cell-cycle checkpoint genes

ABSTRACT

This invention relates to a class of checkpoint genes and their polypeptide products which control progression through the cell cycle in eukaryotic cells. In particular this invention relates to  Schizosaccharomyces pombe  rad3 gene, to its human homologue (ATR) and to their encoded proteins. The invention further relates to assay methods for selecting compounds which modulate the activity of the polypeptide products of these checkpoint genes and the use of the selected compounds in anticancer therapy.

This application claims priority of International Application No: PCT/GB96/09433, filed Sep. 6, 1996, which claims priority of Great Britain Application No: 9518220.0, filed Sep. 6, 1995.

The present invention relates to a class of checkpoint genes which control progression through the cell cycle in eukaryotic cells.

BACKGROUND OF THE INVENTION

Control of the cell cycle is fundamental to the growth and maintenance of eukaryotic organisms, from yeasts to mammals. Eukaryotic cells have evolved control pathways, termed “checkpoints” which ensure that individual steps of the cell cycle are completed before the next step occurs. In response to DNA damage, cell survival is increased both by direct DNA repair mechanisms and by delaying progression through the cell cycle. Depending on the position of the cell within the cycle at the time of irradiation, DNA damage in mammalian cells can prevent (a) passage from G1 into S phase, (b) progression through S phase or (c) passage from G2 into mitosis. Such checkpoints are thought to prevent deleterious events such as replication of damaged DNA and the segregation of fragmented chromosomes during mitosis (Hartwell and Kastan, 1994).

The rad3 gene of Schizosaccharomyces pombe is required for the checkpoints that respond to DNA damage and replication blocks. Rad3 is a member of the lipid kinase subclass of kinases which possess regions having sequence homology to the lipid kinase domain of the p110 subunit of phosphatidylinositol-3 kinase (PI-3 kinase). This subclass also includes the ATM protein defective in ataxia-telangiectasia patients. Cells from aeaxia telangiectasia patients (AT cells) have lost the delay to S phase following irradiation and are said to display radio resistant DNA synthesis (Painter and Young, 1989). AT cells irradiated in S phase accumulate in G2 with lethal damage, presumably as a consequence of attempting to replicate damaged DNA. AT cells irradiated during G2 display a different phenotype, they do not arrest mitosis after DNA damage, and progress through mitosis with damaged DNA (Beamish and Lavin, 1994). Mutations at the A-T locus, to which the ATM gene has been mapped, thus result in disruption of several checkpoints required for an appropriate response to ionising radiation. Other members of this lipid kinase subclass include: Tel1p (Greenwell et al. 1985). a gene involved in maintaining proper telomere length in Saccharomyces cerevisiae; Esr1p; Mec1p and the product of the Drosophila melanogaster mei-41 checkpoint gene (Hari et al. 1995).

DISCLOSURE OF THE INVENTION

We have analyzed the S. pombe rad3 gene and found that it has a full length amino acid sequence of 2386 amino acids, not the 1070 amino acids described by Seaton et al. 1992. We have determined that this is the direct homologue of S. cerevisiae Esr1p, and that it shares the same overall structure as the ATM gene. The C-terminal region of the rad3 protein contains a lipid kinase domain, which is required for Rad3 function We have shown that Rad3 is capable of self association. We have also identified a protein kinase activity associated with Rad3.

Further, we have found a human homologue to rad3. This gene, which we have named ATR (ataxia and rad related), displays significantly higher homology to rad3 than it does to the ATM gene.

The human ATR cDNA sequence is set out as Seq. ID No. 1. The amino acid sequence of the ORF from nucleotides 80 and 8011 is set out as Seq. ID No. 2.

The DNA sequence of the open reading frame (ORF) of rad3 is shown as Seq. ID. No. 3. The 2386 amino acid translation of the gene (nucleotides 585 to 7742 of Seq. ID No. 3) is shown as Seq. ID. No. 4.

Accordingly, in a first aspect, the invention provides the ATR protein of Seq. ID. 2 and homologues thereof, polypeptide fragments thereof, as well as antibodies capable of binding the ATR protein or polypeptide fragments thereof. ATR proteins, homologues and fragments thereof are referred to below as polypeptides of the invention.

In another aspect, the present invention provides a polynucleotide in substantially isolated form capable of hybridising selectively to Seq.ID No. 1 or to the complement (i.e. opposite stand) thereof. Also provided are polynucleotides encoding polypeptides of the invention.

Such polynucleotides will be referred to as a polynucleotide of die invention. A polynucleotides of the invention includes DNA of Seq.ID Nos. 1 and fragments thereof capable of selectively hybridising to this gene.

In a further aspect, the invention provides recombinant vectors carrying a polynucleotide of the invention, including expression vectors, and methods of growing such vectors in a suitable host cell, for example under conditions in which expression of a protein or polypeptide encoded by a sequence of the invention occurs.

In an additional aspect, the invention provides kits comprising polynucleotides, polypeptides or antibodies of the invention and methods of using such kits in diagnosing the presence of absence of ATR and its homologues, or variants thereof, including deleterious ATR mutants.

The invention further provides assay methods for screening candidate substances for use as compounds for inhibiting or activating ATR activity, or the activity of mutated forms of ATR which are deficient in checkpoint activity. The invention also provides assay methods for screening candidate substances for use as compounds for inhibiting interactions between ATR and other compounds that interact with ATR, including ATR itself.

In a related aspect, the invention also provides a polynucleotide sequence of Seq. ID No. 3 in substantially isolated form, and the protein of Seq. ID No. 4 in substantially isolated form, and novel fragments and variants thereof.

DETAILED DESCRIPTION OF THE INVENTION

A. Polynucleotides

Polynucleotides of the invention may comprise DNA or RNA. They may also be polynucleotides which include within them synthetic or modified nucleotides. A number of different types of modification to oligonucleotides are known in the an. These include methylphosphonate and phosphorothioate backbones, addition of acridine or polylysine chains at the 3′ and/or 5′ ends of the molecule. For the purposes of the present invention, it is to be understood that the polynucleotides described herein may be modified by any method available in the art. Such modifications may be carried out in order to enhance the in vivo activity or lifespan of polynucleotides of the invention.

Polynucleotides of the invention capable of selectively hybridizing to the DNA of Seq. ID No. 1 will be generally at least 70%, preferably at least 80 or 90% and more preferably at least 95% homologous to the corresponding DNA of Seq. ID No. 1 over a region of at least 20, preferably at least 25 or 30, for instance at least 40, 60 or 100 or more contiguous nucleotides.

It is to be understood that skilled persons may, using routine techniques, make nucleotide substitutions that do not affect the polypeptide sequence encoded by the polynucleotides of the invention to reflect the codon usage of any particular host organism in which the polypeptides of the invention are to be expressed.

Any combination of the above mentioned degrees of homology and minimum sizes may be used to define polynucleotides of the invention, with the more stringent combinations (i.e. higher homology over longer lengths) being preferred. Thus for example a polynucleotide which is at least 80% homologous over 25, preferably 30 nucleotides forms one aspect of the invention, as does a polynucleotide which is at least 90% homologous over 40 nucleotides.

Polynucleotides of the invention may be used to produce a primer, e.g. a PCR primer, a primer for an alternative amplification reaction, a probe e.g. labelled with a revealing label by conventional means using radioactive or non-radioactive labels, or the polynucleotides may be cloned into vectors. Such primers, probes and other fragments will be at least 15, preferably at least 20, for example at least 25, 30 or 40 nucleotides in length, and are also encompassed by the term polynucleotides of the invention as used herein.

Polynucleotides such as a DNA polynucleotide and primers according to the invention may be produced recombinantly, synthetically, or by any means available to those of skill in the art. They may also be cloned by standard techniques.

In general, primers will be produced by synthetic means, involving a step wise manufacture of the desired nucleic acid sequence one nucleotide at a time. Techniques for accomplishing this using automated techniques are readily available in the art.

Longer polynucleotides will generally be produced using recombinant means, for example using a PCR (polymerase chain reaction) cloning techniques. This will involve making a pair of primers (e.g. of about 15-30 nucleotides) to a region of the ATR gene which it is desired to clone, bringing the primers into contact with mRNA or cDNA obtained from a human cell (e.g. a dividing cell such as a peripheral blood leukocyte), performing a polymerase chain reaction under conditions which bring about amplification of the desired region, isolating the amplified fragment (e.g. by purifying the reaction mixture on an agarose gel) and recovering the amplified DNA. The primers may be designed to contain suitable restriction enzyme recognition sites so that be amplified DNA can be cloned into a suitable cloning vector.

Such techniques may be used to obtain all or part of the ATR sequence described herein. Genomic clones containing the ATR gene and its introns and promoter regions may also be obtained in an analogous manner, staring with genomic DNA from a human cell, e.g. a liver cell.

Although in general the techniques mentioned herein are well known in the art, reference may be made in particular to Sambrook et al. (Molecular Cloning: A Laboratory Manual, 1989).

Polynucleotides which are not 100% homologous to the sequences of the present invention but fall within the scope of the invention can be obtained in a number of ways.

Other human allelic variants of the ATR sequence described herein may be obtained for example by probing genomic DNA libraries made from a range of individuals, for example individuals from different populations.

In addition, other animal, particularly mammalian (e.g. mice, rats or rabbits), more particularly pa, homologues of ATR may be obtained and such homologues and fragments thereof in general will be capable of selectively hybridizing to Seq. ID No. 1. Such sequences may be obtained by probing cDNA libraries made from dividing cells or tissues or genomic DNA libraries from other animal species, and probing such libraries with probes comprising all or part of Seq. ID. 1 under conditions of medium to high stringency (for example 0.03M sodium chloride and 0.03M sodium citrate at from about 50° C. to about 60° C.).

Allelic variants and species homologues may also be obtained using degenerate PCR which will use primers designed to target sequences within the variants and homologues encoding conserved amino acid sequences. Conserved sequences can be predicted from aligning the AIR amino acid sequence with that of rad3. The primers will contain one or more degenerate positions and will be used at stringency conditions lower than those used for cloning sequences with single sequence primers against known sequences.

Alternatively, such polynucleotides may be obtained by site directed mutagenesis of the ATR sequences or allelic variants thereof. This may be useful where for example silent codon changes are required to sequences to optimise codon preferences for a particular host cell in which the polynucleotide sequences are being expressed. Other sequence changes may be desired in order to introduce restriction enzyme recognition sites, or to alter the property or function of the polypeptides encoded by the polynucleotides. Further changes may be desirable to represent particular coding changes found in ATR which give rise to mutant ATR genes which have lost the checkpoint fiction. Probes based on such changes can be used as diagnostic probes to detect such ATR mutants.

The invention further provides double stranded polynucleotides comprising a polynucleotide of the invention and its complement.

Polynucleotides or primers of the invention may carry a revealing label. Suitable labels include radioisotopes such as ³²P or ³⁵S, enzyme labels, or other protein labels such as biotin. Such labels may be added to polynucleotides or primers of the invention and may be detected using by techniques known per se.

Polynucleotides or primers of the invention or fragments thereof labelled or unlabelled may be used by a person skilled in the art in nucleic acid-based tests for detecting or sequencing ATR in the human or animal body.

Such tests for detecting generally comprise bringing a human or animal body sample contain DNA or RNA into contact with a probe comprising a polynucleotide or primer of the invention under hybridizing conditions and detecting any duplex formed between the probe and nucleic acid in the sample. Such detection may be achieved using techniques such as PCR or by immobilizing the probe on a solid support, removing nucleic acid in the sample which is not hybridized to the probe, and then detecting nucleic acid which has hybridized to the probe. Alternatively, the sample nucleic acid may be immobilized on a solid support, and the amount of probe bound to such a support can be detected. Suitable assay methods of this any other formats can be found in for example WO89/03891 and WO90/13667.

Tests for sequencing ATR include bringing a human or animal body sample containing target DNA or RNA into contact with a probe comprising a polynucleotide or primer of the invention under hybridizing conditions and determining the she by, for example the Sanger dideoxy chain termination method (see Sambrook et al.).

Such a method generally comprises elongating, in the presence of suitable reagents, the primer by synthesis of a stand complementary to the target DNA or RNA and selectively terminating the elongation reaction at one or more of an A, C, G or T/U residue; allowing stand elongation and termination reaction to occur; separating out according to size the elongated products to determine the sequence of the nucleotides at which selective tenon has occurred. Suitable reagents include a DNA polymerase enzyme, the deoxynucleotides dATP, dCTP, dGTP and dTTP, a buffer and ATP. Dideoxynucleotides are used for selective termination.

Tests for detecting or sequencing ATR in the human or animal body may be used to determine ATR sequences within cells in individuals who have, or are suspected to have, an altered ATR gene sequence, for example within cancer cells including leukaemic cells and solid tumours such as breast, ovary, lung, colon, pancreas, testes, liver, brain, muscle and bone tumours.

In addition, the discovery of ATR will allow the role of this gene in hereditary diseases to be investigated, in a manner analogous to the ATM gene. In general, this will involve establishing the status of ATR (e.g using PCR sequence analysis) in cells derived from patients with diseases that may be connected with damage to replicating cells, e.g. familial predisposition to cancer. chromosome breakage or instability phenotype or repair-damage sensitivity phenotype.

The probes of the invention may conveniently be packaged in the form of a test kit in a suitable container. In such kits the probe may be bound to a solid support where the assay format for which the kit is designed requires such binding. The kit may also contain suitable reagents for treating the sample to be probed, hybridizing the probe to nucleic acid in the sample, control reagents, instructions, and the like.

The present invention also provides polynucleotides encoding the polypeptides of the invention described below. Because such polynucleotides will be useful as sequences for recombinant production of polypeptides of the invention, it is not necessary for them to be selectively hybridizable to the sequence Seq. ID No. 1, although this will generally be desirable. Otherwise, such polynucleotides may be labelled, used, and made as described above if desired. Polypeptides of the invention are described below.

Particularly preferred polynucleotides of the invention are hose derived from the lipid kinase domain of ATR, its allelic variants and species homologues. The lipid kinase domain is represented by nucleotides 7054 to 8011 of Seq. ID. 1. Polynucleotides of the invention which comprise this domain are particularly preferred. The “lipid kinase domain” refers to a domain which has homology to other known lipid kinases, in particular the p110 subunit of PI-3 kinase, as determined by sequence alignments.

Other preferred polynucleotides of the invention those which comprise nucleotides encoding amino acids 181 to 302 of Seq. ID No. 2 (nucleotides 620 to 985 of Seq. ID No. 1), which is believed to be a leucine zipper region, a putative site of protein-protein interaction, and amino acids 1358 to 1366 (nucleotides 4151 to 4177), which is also conserved. In an additional aspect, polynucleotides of the invention include those of Seq. ID No. 3 and fragments thereof capable of selectively hybridizing to this sequence other than the fragment consisting of nucleotides 2482 to 6599 in which the following changes have been made: Deletion of residues 2499, 2501, 2507 & 2509; insertion of C between 5918/5919.

Particularly preferred fragments include those comprising residues 6826 to 7334 (the lipid kinase domain) and the leucine zipper regions 1476 to 1625 and 2310 to 2357. Additionally, the fragment comprising the conserved region 3891 to 3917 is preferred. Such polypeptides and fragments may be made and used as described above.

B. Polypeptides

Polypeptides of the invention include polypeptides in substantially isolated form which comprise the sequence set out in Seq ID No. 2.

Polypeptides further include van of such sequences, including naturally occurring allelic variants and synthetic variants which are substantially homologous to said polypeptides. In this context, substantial homology is regarded as a sequence which has at least 70%, e.g. 80% or 90% amino acid homology (identity) over 30 amino acids with the sequence of Seq. ID No. 2 except for the lipid kinase domain and C-terminal portion (residues 2326 to 2644) where substantial homology is regarded as at least 80% homology, preferably 90% homology (identity) over 50 amino acids.

Polypeptides also include other those encoding ATR homologues from other species including animals such as mammals (e.g. mice, rats or rabbits), especially primates, and variants thereof as defined above.

Polypeptides of the invention also include fragments of the above mentioned full length polypeptides and variants thereof, including fragments of the sequence set out in Seq. ID No. 2.

Preferred fragments include those which include an epitope, especially an epitope. Suitable fragments will be at least about 5, e.g. 10, 12, 15 or 20 amino acids in size. Polypeptide fragments of the ATR protein and allelic and species variants thereof may contain one or more (e.g. 2, 3, 5, or 10) substitutions, deletions or insertions, including conserved substitutions.

Conserved substitutions may be made according to the following table indicates conservative substitutions, where amino acids on the same block in the second column and preferably in the same line in the third column may be substituted for each other:

ALIPHATIC Non-polar G A P I L V Polar - uncharged C S T M N Q Polar - charged D E K R AROMATIC H F W Y OTHER N Q D E

Variants of the polypeptides of the invention may also comprise polypeptides wherein one or more of the specified (i.e., naturally encoded) amino acids is deleted or replaced or wherein one or more nonspecified ammo acids arc added: (1) without loss of the kinase activity specific to the polypeptides of the invention; or (2) with disablement of the kinase activity specific to the polypeptides of the invention; or (3) with disablement of the ability to intend with members or regulators of the cell cycle checkpoint pathway.

Epitopes may be determined either by techniques such as peptide scanning techniques as described by Geysen et al. Mol. Immunol., 23; 709-715 (1986).

Polypeptides of the invention may be in a substantially isolated form. It will be understood that the polypeptide may be mixed with carriers or diluents which will not interfere with the intended purpose of the polypeptide and still be regarded as substantially isolated. A polypeptide of the invention may also be in a substantially purified form, in which case it will generally comprise the polypeptide in a preparation in which more than 90%, e.g. 95%, 98% or 99% of the polypeptide in the preparation is a polypeptide of the invention. Polypeptides of the invention may be modified for example by the addition of Histidine residues to assist their purification or by the addition of a signal sequence to promote their secretion from a cell.

A polypeptide of the invention may be labelled with a revealing label. The revealing label may be any suitable label which allows the polypeptide to be detected. Suitable labels include radioisotopes, e.g. ¹²⁵I, enzymes, antibodies, polynucleotides and linkers such as biotin. Labelled polypeptides of the invention may be used in diagnostic procedures such as immunoassays in order to determine the amount of a polypeptide of the invention in a sample. Polypeptides or labelled polypeptides of the invention may also be used in serological or cell mediated immune assays for the detection of immune reactivity to said polypeptides in animals and humans using standard protocols.

A polypeptide or labelled polypeptide of the invention or fragment thereof may also be fixed to a solid phase, for example the surface of an immunoassay well or dipstick.

Such labelled and/or immobilized polypeptides may be paged into kits in a suitable container along with suitable reagents, controls, instructions and the like.

Such polypeptides and kits may be used in methods of detection of antibodies to the ATR protein or its allelic or species variants by immunoassay.

Immunoassay methods are well known in the an and will generally comprise:

(a) providing a polypeptide comprising an epitope bindable by an antibody against said protein;

(b) incubating a biological sample with said polypeptide under conditions which allow for the formation of an antibody-antigen complex; and

(c) determining whether antibody-antigen complex comprising said polypeptide is formed.

Polypeptides of the invention may be may by synthetic means (e.g. as described by Geysen et al.) or recombinantly, as described below.

Particularly preferred polypeptides of the invention include those spanning or within the lipid kinase domain, namely from amino acids 2326 to 2644 of Seq. ID. 2. or sequences substantially homologous thereto. Fragments as defined above from this region are particularly preferred. The polypeptides and fragments thereof may contain amino acid alterations as defined above, including substitutions at one or more of positions 2475, 2480 and 2494, which correspond to the positions of the rad3 substitutions described in the examples below. Preferred substiutions include D2475A, N2480K and D2494E.

Polypeptides of the invention may be used in in vitro or in vivo cell culture systems to study the role of ATR as a checkpoint gene. For example, truncated or modified (e.g. modified in the lipid kinase domain) ATRs may be introduced into a cell to disrupt the normal checkpoint functions which occur in the cell.

The polypeptides of the invention may be introduced into the cell by in situ expression of the polypeptide from a recombinant expression vector (see below). The expression vector optionally carries an inducible promoter to control the expression of the polypeptide.

The use of mammalian host cells is expected to provide for such post-translational modifications (e.g., myristolation, glycosylation, truncation, lapidation and tyrosine, serine or threonine phosphorylation) as may be needed to confer optimal biological activity on recombinant expression products of the invention.

Such cell culture systems in which polypeptide of the invention are expressed may be used in assay systems to identify candidate substances which interfere or enhance checkpoint functions in the cell (see below).

In an additional aspect, polypeptides of the invention include the protein of Seq. ID No. 4 and fragments thereof from the region other than the fragment consisting of amino acids 713 to 1778. Particularly preferred fragments include those comprising residues 2082 to 2386 (the lipid kinase domain) and the leucine zipper regions 298 to 347 and 576 to 591. Additionally, the fragment comprising the conserved region 1103 to 1111 is preferred. Such polypeptides and fragments may be made and used as described above.

The invention also provides polypeptides substantially homologous to the protein of Seq. ID No. 4, and fragments thereof. In this context, substantial homology is regarded as a sequence which has at least 70%, e.g. 80% or 90% amino acid homology (identity) over 30 amino acids with the sequence of Seq. ID No. 4 except for the lipid kinase domain and C-terminal portion (residues 2082 to 2386) where substantial homology is regarded as at least 80%, preferably at least 90% homology (identity) over 50 amino acids.

C. Vectors

Polynucleotides of the invention can be incorporated into a recombinant replicable vector. The vector may be used to replicate the nucleic acid in a compatible host cell. Thus in a further embodiment, the invention provides a method of making polynucleotide of the invention by introducing a polynucleotide of the invention into a replicable vector, introducing the vector into a compatible host cell, and growing the host cell under conditions which bring about replication of the vector. The vector may be recovered from the host cell. Suitable host cells are described below in connection with expression vectors.

D. Expression Vectors

Preferably, a polynucleotide of the invention in a vector is operably linked to a control sequence which is capable of providing for the expression of the coding sequence by the host cell, i.e. the vector is an expression vector.

The term “operably linked” refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner. A control sequence “operably linked” to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under condition compatible with the control sequences.

Such vectors may be transformed into a suitable host cell as described above to provide for expression of a polypeptide of the invention. Thus, in a further aspect the invention provides a process for preparing polypeptides according to the invention which comprises cultivating a host cell transformed or transfected with an expression vector as described above under conditions to provide for expression by the vector of a coding sequence encoding the polypeptides, and recovering the expressed polypeptides.

The vectors may be for example, plasmid, virus or phage vectors provided with an origin of replication, optionally a promoter for the expression of the said polynucleotide and optionally a regulator of the promoter. The vectors may contain one or more selectable marker genes, for example an ampicillin resistance gene in the case of a bacterial plasmid or a neomycin resistance gene for a mammalian vector. Vectors may be used in vitro, for example for the production of RNA or used to transfect or transform a host cell. The vector may also be adapted to be used in vivo, for example in a method of gene therapy.

A further embodiment of the invention provides host cells transformed or transfected with the vectors for the replication and expression of polynucleotides of the invention. The cells will be chosen to be compatible with the said vector and may for example be bacterial, yeast, insect or mammalian.

Polynucleotides according to the invention may also be inserted into the vectors described above in an antisense orientation in order to provide for the production of antisense RNA. Antisense RNA or other antisense polynucleotides may also be produced by synthetic means. Such antisense polynucleotides may be used in a method of controlling the levels of ATR or its variants or species homologues.

Promoters and other expression regulation signals may be selected to be compatible with the host cell for which the expression vector is designed. For example, yeast promoters include S. cerevisiae GAL4 and ADH promoters, S. pombe nmt1 and adh promoter. Mammalian promoters include the metallothionein promoter which is can be included in response to heavy metals such as cadmium. Viral promoters such as the SV40 large T antigen promoter or adenovirus promoters may also be used. All these promoters are readily available in the art.

E. Antibodies

The invention also provides monoclonal or polyclonal antibodies to polypeptides of the invention or fragments thereof. The invention further provides a process for the production of monoclonal or polyclonal antibodies to polypeptides of the invention. Monoclonal antibodies may be prepared by conventional hybridoma technology using the polypeptides of the invention or peptide fragments thereof, as immunogens. Polyclonal antibodies may also be prepared by conventional means which comprise inoculating a host animal, for example a rat or a rabbit, with a polypeptide of the invention or peptide fragment thereof and recovering immune serum.

In order that such antibodies may be made, the invention also provides polypeptides of the invention or fragments thereof haptenised to another polypeptide for use as immunogens in animals or humans

Preferred antibodies of the invention will be capable of selectively binding the human ATR protein, that is with an affinity at least 10 fold, preferably at least 100 fold that of the rad3 protein. Such antibodies can be obtained by routine experimentation, e.g. selecting regions of ATR protein with sequences different from the corresponding regions of rad3, making peptides comprising such sequences and using such peptides as immunogens. Following production of antibodies the binding of said antibodies may be determined. Preferred antibodies of the invention include those capable of selectively binding the lipid kinase domain (as defined above) of the human ATR protein. In addition, antibodies which are capable of binding the human and yeast (S. pombe) lipid kinase domains with similar affinity, but not to the domains of the ATM family of proteins form a further aspect of the invention. Such antibodies may be raised against peptides from the lipid se domains which correspond to regions found to be identical, or substantially identical, in the yeast and human genes.

For the purposes of this invention, the term “antibody”, unless specified to the contrary, includes fragments of whole antibodies which retain their binding activity for a tumour target antigen. Such fragments include Fv, F(ab′) and F(ab′)₂ fragments, as well as single chain antibodies. Furthermore, the antibodies and fragments thereof may be humanised antibodies, eg. as described in EP-A-239400.

Antibodies may be used in method of detecting polypeptides of the invention present in biological samples by a method which comprises:

(a) providing an antibody of the invention;

(b) incubating a biological sample with said antibody under conditions which allow for the formation of an antibody-antigen complex; and

(c) determining whether antibody-antigen complex comprising said antibody is formed.

Suitable samples include extracts from dividing cells, e.g leukocytes or cancer cells including leukaemic cells and solid tumours such as breast, ovary, lung, colon, pancreas, testes, liver, brain, muscle and bone tumours.

Antibodies of the invention may be bound to a solid support and/or packaged into kits in a suitable container along with suitable reagents, controls, instructions and die like.

F. Assays

Abrogating cell cycle checkpoints is a potential strategy for developing or designing drugs for anti cancer therapy, both as a novel treatment as such and as part of a combination therapy to enhance the specific toxicity of current chemotherapeutic agents. For example alkylating agents such as nitrogen mustards are used a chemotherapeutic agents which damage DNA in rapidly dividing cells, leading to cell death. The toxicity of such agents may be lessened by DNA repair and checkpoint mechanisms. Abrogating such mechanisms will thus enhance the effectiveness of therapeutic compounds designed to damage DNA. Abrogation of the ATR checkpoint will be especially useful where tumour cells have lost other checkpoint or damage response genes, since these other genes may be able to complement the loss of ATR function in non tumour cells, leading to an even greater enhancement in the effectiveness of the chemotherapeutic agent.

The lipid kinase activity of ATR is a target for developing anticancer compounds, since the results presented in the following examples indicate that the kinase domain is required for ATR function. Thus the present invention provides an assay method for screening candidate substances for anti-cancer therapy which comprises:

(a) providing a polypeptide of the invention which retains lipid kinase activity and a substrate for said kinase, under conditions and with reagents such that the kinase activity will act upon the substrate;

(b) bringing said polypeptide and substrate into contact with a candidate substance;

(c) measuring the degree of decrease in the kinase activity of the polypeptide; and

(d) selecting a candidate substance which provides a decrease in activity.

The assay may be carried out in vitro, for example in the wells of a microtitre dish. Such a format may be readily adapted for automation, allowing large numbers of candidate substances to be screened.

The substrate may be a protein or lipid substrate of natural or synthetic origin upon which the polypeptide of the invention will act. Usually, the polypeptide of the invention will phosphorylate the substrate.

Any suitable format for the assay may be used by those of skill in the art of throughput assays. Typically, the polypeptide of the invention which retains lipid kinase activity will be bound to a solid support in the presence of a substrate and cellular and other components which are usually required for activity. Labelled phosphate and a candidate substance will be added to the mixture simultaneously or sequentially in either order. After a suitable reaction time (usually a few minutes but in any event enough for phosphorylation of the substrate in the absence of candidate substance to occur) the amount of free phosphate is determined, e.g. by precipitation of phosphate. Candidate substances which inhibit kinase activity will inhibit the incorporation of free phosphate into the substrate and thus where free phosphate is found this is indicative of inhibition.

Other assay formats may be used by those skilled in the art.

The candidate substances may be used in an initial screen in batches of for example 10 compounds per reaction, and the compounds of those batches which show inhibition tested individually.

Suitable candidate substances include peptides, especially of from about 5 to 20 amino acids in size, based on the sequence of the kinase domain, or variants of such peptides in which one or more residues have been substituted as described above. Peptides from panels of peptides comprising random sequences or sequences which have been varied consistently to provide a maximally diverse panel of peptides may be used. Further candidate substances include kinase inhibitors which are small molecules such as cyclosporin-like and staurosporin-like compounds, or other compounds commercially available in panels of small molecule inhibitors.

Candidate substances which show activity in in vitro screens such as the above can then be tested in in vivo systems, such as yeast or mammalian cells which will be exposed to the inhibitor and tested for checkpoint activity.

We have also shown that Rad3 possesses protein kinase activity. Target substrates of Rad3 protein kinase activity may be identified by incorporating test compounds in assays for kinase activity. Rad3 protein is resuspended in kinase buffer and incubated either in the presence of absence of the test compound (e.g., casein, histone H1, or appropriate substrate peptide). Moles of phosphate transferred by the kinase to the test compound are measured by autoradiography or scintillation courting. Transfer of phosphate to the test compound is indicative that the test compound is a substrate of the kinase.

Agents that modulate Rad3/ATR lipid kinase or Rad 3 protein kinase activity may be identified by incubating a test compound and Rad3/ATR immunopurified from cells naturally expressing Rad3/ATR, with Rad3/ATR obtained from recombinant procaryotic or eukaryotic cells expressing the enzyme, or with purified Rad3/ATR, and then determining the effect of the test compound on Rad3/ATR activity. The activity of the Rad3/ATR lipid kinase or Rad3 protein kinase domains can be measured by determining the moles of ³²P-phosphate transferred by the kinase from gamma-³²-P-ATP to either itself (autophosphorylation) or to an exogenous substrate such as a lipid or protein. The amount of phosphate incorporated into the substrate is measured by scintillation counting or autoradiography. An increase in the moles of phosphate transferred to the substrate in the presence of the test compound compared to the moles of phosphate transferred to the substrate in the absence of the test compound indicates that the test compound is an activator of said kinase activity. Conversely, a decrease in the moles of phosphate transferred to the substrate in presence of the test compound compared to the moles of phosphate transferred to the substrate in the absence of the test compound indicates that the modulator is an inhibitor of said kinase activity.

In a presently preferred assay, a Rad3/ATR antibody linked to agarose beads is incubated with a cell lysate prepared from host cells expressing Rad3/ATR. The beads are washed to remove proteins binding nonspecifically to the beads and the beads are then resuspended in a kinase buffer (such as 25 mM K-HEPES pH 7.7, 50 mM potassium chloride, 10 mM magnesium chloride. 0.1% Nonidet-P40, 20% glycerol, 1 mM DTT). The reaction is initiated by the addition of 100 μM gamma-³²P-ATP (4 Ci/mM) and an exogenous substrate such as lipid or peptide, and the reaction is carried out at 30° C. for 10 minutes. The activity of the kinase is measured by determining the moles of ³²P-phosphate transferred either to the kinase itself or the added substrate. In a preferred embodiment the host cells lack endogenous Rad3/ATR kinase activity. The selectivity of a compound that modulates the lipid kinase activity of Rad3/ATR can be evaluated by comparing its activity on Rad3/ATR to its activity on, for example, other known phosphatidylinositol-3 (PI-3)related kinases. The combination of the recombinant Rad3/ATR products of the invention with other recombinant PI-3-related kinase products in a series of independent assays provides a system for developing selective modulators of Rad3/ATR kinase activity. Similarly, the selectivity of a compound that modulates the protein kinase activity of Rad3 may be determined with reference to other protein kinases, for example the DNA dependent protein kinase or ATM.

In addition, the demonstration that the rad mutant rad.D2249E (see Examples) can act as a dominant negative mutant indicates involvement in one or more protein complexes, and such complexes themselves can be targeted for therapeutic interventions We have shown, for example, that Rad3 can both self associate and associate with ATR. It is therefore likely that Rad/ATR function as multimeric molecules Mutant yeast rad or human ATR genes, or derivatives thereof which also lack rad/ATR activity may be introduced into cells to act as dominant negative mutants. Thus for example if expression of a dominant negative mutant (e.g. ATR D2475A, N2480K or D2494E) in a tumour cell leads to enhanced radiation sensitivity this indicates that the native ATR is still functioning and thus a target for therapeutic agents.

Interacting proteins including components of multimeric protein complexes involving Rad3 or ATR may be identified by the following assays.

A first assay contemplated by the invention is a two-hybrid screen. The two-hybrid system was developed in yeast (Chien et al. (1991)) and is based on functional in vivo reconstitution of a transcription factor which activates a reporter gene. Specifically, a polynucleotide encoding a protein that interacts with Rad3/ATR is isolated by: transforming or transfecting appropriate host cells with a DNA construct comprising a reporter gene under the control of a promoter regulated by a transcription factor having DNA a binding domain and an activating domain: expressing in the host cells a first hybrid DNA sequence encoding a fire fusion of part or all of Rad3/ATR and either the DNA binding domain or the activating domain of the transcription factor; expressing in the host cell a library of second hybrid DNA sequences encoding second fusion of part or all putative Rad3/ATR binding proteins and the DNA binding domain or activating domain of the transcription factor which is not incorporated in the first fusion; detecting binding of an Rad3/ATR interacting protein to Rad3/ATR in a particular host cell by detecting the production of reporter gene product in the host cell; and isolating second hybrid DNA sequences encoding the interacting protein from the particular host cell. Presently preferred for use in the assay are a lexA promoter to drive expression of the reporter gene, the reporter gene, a transcription factor comprising the lexA DNA binding domain and the GAL4 transactivation domain, and yeast host cells.

Other assays for identifying proteins that interact with Rad3 or ATR may involve immobilising Rad3/ATR or a test protein, detectably labelling the nonimmobilised binding partner, incubating the binding partners together and determining the amount of label bound. Bound label indicates that the test protein interacts with Rad3/ATR.

Another type of assay for identifying Rad3 or ATR interacting proteins involves immobilising Rad3/ATR or a fragment thereof on a solid support coated (or impregnated with) a fluorescent agent, labelling a test protein with a compound capable of exciting the fluorescent agent, contacting the immobilised Rad3/ATR with the labelled test protein, detecting light emission by the fluorescent agent, and identifying interacting proteins as test proteins which result in the emission of light by the fluorescent agent. Alternatively, the putative interacting protein may be immobilised and Rad3/ATR may be labelled in the assay.

Compounds that modulate interaction between Rad3/ATR and other cellular components may be used in methods of treating cancer. For example, if a particular form of cancer results from a mutation in a gene other than ATR such as the p53 gene, an agent which inhibits the transcription or the enzymatic activity of ATR and thus the G₂ cell cycle checkpoint may be used to render cancerous cells more susceptible to chemotherapy or radiation therapy. The therapeutic value of such an agent lies in the fact that current radiation therapy or chemotherapy in most cases does nothing to overcome the ability of the p53 mutant cancerous cell to sense and correct the DNA damage imposed as a result of the treatment. As a result, a cancer cell can simply repair the DNA damage. Modulating agents of the invention may therefore be chemotherapy and radiation adjuvants or may be directly active as chemotherapy drugs themselves.

Assays for identifying compounds that modulate interaction of Rad3/ATR with other proteins may involve: transforming or transfecting appropriate host cells with a DNA construct comprising a reporter gene under the control of a promoter regulated by a transcription factor having a DNA-binding domain and an activating domain: expressing in the host cells a first hybrid DNA sequence encoding a first fusion of part or all of Rad3/ATR and the DNA binding domain or the activating domain of the transcription factor; expressing in the host cells a second hybrid DNA sequence encoding part or all of a protein that interacts with Rad3/ATR and the DNA binding domain or activating domain of the transcription factor which is not incorporated in the first fusion; evaluating the effect of a test compound on the interaction between Rad3/ATR and the interacting protein by detecting binding of the interacting protein to Rad3/ATR in a particular host cell by measuring the production of reporter gene product in the host cell in the presence or absence of the test compound; and identifying modulating compounds as those test compounds altering production of the reported gene product in comparison to production of the reporter gene product in the absence of the modulating compound. Presently preferred for use in the assay are a lexA promoter to drive expression of the reporter gene, the lacZ reporter gene, a transcription factor comprising the lexA DNA domain and the GALA transactivation domain, and yeast host cells.

Another type of assay for identifying compounds that modulate the interaction between Rad3/ATR and an interacting protein involves immobilising Rad3/ATR or a natural Rad3/ATR interacting protein, detectably labelling the nonimmobilised binding partner, incubating the binding partners together and determining the effect of a test compound on the amount of label bound wherein a reduction in the label bound in the present of the test compound compared to the amount of label bound in the absence of the test compound indicates that the test agent is an inhibitor of Rad3/ATR interaction with the protein. Conversely, an increase in the bound in the presence of the test compared to the amount label bound in the absence of the compared indicates that the putative modulator is an activator of Rad3/ATR interaction with the protein.

Yet another method contemplated by the invention for identifying compounds that modulate the binding between Rad3/ATR and an interacting protein involves immobilising Rad3/ATR or a fragment thereof on a solid support coated (or impregnated with) a fluorescent agent, labelling the interacting protein with a compound capable of exciting the fluorescent agent, contacting the immobilised Rad3/ATR with the labelled interacting protein in the presence and absence of a test compound, detecting light emission by the fluorescent agent, and identifying modulating compounds as those test compounds that affect the emission of light by the fluorescent agent in comparison to the emission of light by the fluorescent agent in the absence of the test compound. Alternatively, the Rad3/ATR interacting protein may be immobilised and Rad3/ATR may be labelled in the assay.

We have shown that Rad3 interacts with ATR. Therefore the above-mentioned assays may also be used to identify compounds that modulate the interaction between Rad3 and ATR where the interacting protein described in the assay methods is either Rad3 or ATR.

We have also shown that Rad3 can bind to itself, strongly suggesting that ATR can also bind to itself. Therefore the above-mentioned assays may also be used to identify compounds that modulate Rad3-Rad3 interactions and ATR-ATR interactions.

Such compounds could be used therapeutically to disrupt ATR-ATR interactions and increase the sensitivity of tumour cells to chemotherapy and/or radiotherapy. Thus the invention provides an assay method for screening candidate substances for anti cancer therapy which comprises:

(a) (i) incubating a polypeptide of the invention with another polypeptide of the invention, which may be the same as or different to the first polypeptide, under conditions which allow the first polypeptide to bind to the second polypeptide to form a complex;

(ii) bringing the complex thus formed into contact with a candidate substance;

or

(a) incubating a polypeptide of the invention with another polypeptide of the invention, which may be the same as or different to the first polypeptide, under conditions which allow the first polypeptide to bind to the second polypeptide to form a complex and in the presence of a candidate substance;

and

(b) determining whether the candidate substance inhibits binding of the first polypeptide to the second polypeptide and

(c) selecting a candidate substance which inhibits binding of the first polypeptide to the second polypeptide.

Preferably the first and second polypeptide may be distinguished from each other. For example, the first polypeptide and the second polypeptide may both be ATR, or may both be Rad3, or one may be ATR and one may be Rad3 or derivatives of either ATR or Rad3 which retain binding activity. When both polypeptides are ATR or Rad3, preferably two distinguishable forms of ATR/Rad3 would be used in these assays. They may be distinguished by, for example, labelling either of the polypeptides. Examples of labels include radioactive labels. epitope tags or other polypeptide tags such as glutathione-S-transferase. For example, one form of Rad3 may have one form of epitope tag, and the other form would have a different epitope tag, allowing them to be distinguished immunologically such that binding of one to the other can be ascertained quantitively or qualitatively. In a preferred method, the first polypeptide may be immobilised, for example to agarose beads or a solid support, and the second polypeptide may be in free solution. Binding is then determined using methods described above and well-known to skilled persons.

Also comprehended by the present invention are antibody products (e.g., monoclonal and polyclonal antibodies, single chain antibodies, chimeric antibodies, CDR-grafted antibodies and the like) and other binding proteins (such as those identified in the assays above) which are specific for the Rad3 protein kinase domain or the Rad3/ATR lipid kinase domains. Binding proteins can be developed using isolated natural or recombinant enzymes. The binding proteins are useful, in turn, for purifying recombinant and naturally occurring enzymes and identifying cells producing such enzymes. Assays for the detection and quantification of proteins in cells and in fluids may involve a single antibody substance or multiple antibody substances in a “sandwich” assay format. The binding proteins are also manifestly useful in modulating (i.e., blocking, inhibiting, or stimulating) enzyme/substrate or enzyme/regulator interactions.

Modulators of Rad3/ATR may affect its kinase activity, its localisation in the cell, and/or its interaction with members of the cell cycle checkpoint pathway. Selective modulators may include, for example, polypeptides or peptides which specifically bind to Rad3/ATR or Rad3/ATR nucleic acid, and/or other non-peptide compounds (e.g., isolated or synthetic organic molecules) which specifically react with Rad3/ATR or Rad3/ATR nucleic acid. Mutant forms of Rad3/ATR which affect the enzymatic activity or cellular localisation of wild-type Rad3/ATR are also contemplated by the invention.

Furthermore, combinatorial libraries, peptide and peptide mimetics, defined chemical entities, oligonucleotides, and natural product libraries may be screened for activity as modulators of Rad3/ATR kinase activity and RAd3/ATR interactions in assays such as those described

F. Therapeutic Uses

Modulators of Rad3/ATR activity, including inhibitors of their lipid kinase and protein kinase activities, may be used in anti-cancer therapy. In particular, they may be used to increase the susceptibility of cancer cells to chemotherapy and/or radiotherapy by virtue of their ability to disrupt the cell cycle regulatory functions of Rad3/ATR.

Thus the invention provides the use of compounds that modulate Rad3/ATR activity, identified by the screening assays described above, in a method of treatment of cancer. In one embodiment, said compounds are capable of inhibiting rad3/ATR lipid kinase and/or Rad3 protein kinase activity. In another embodiment, said compounds are capable of inhibiting interactions between ATR and itself and/or between ATR and other interacting proteins which may, for example, normally form part of a multimeric protein complex.

It is to be understood that the term “compound” in this context also refers 10 the candidate substances selected in the above-described assays.

Typically the compounds are formulated for clinical administration by mixing them with a pharmaceutically acceptable carrier or diluent. For example they can be formulated for topical, parenteral, intravenous, intramuscular, subcutaneous, intraocular or transdermal administration. Preferably, the compound is used in an injectable form. Direct injection into the patient's tumour is advantageous because it makes it possible to concentrate the therapeutic effect at the level of the affected tissues. It may therefore be so with any vehicle which is pharmaceutically acceptable for an injectable formulation, preferably for a direct injection at the site to be treated. The pharmaceutically carrier or diluent may be, for example, sterile or isotonic solutions.

The dose of compound used may be adjusted according to various parameters, especially according to the compound used, the age, weight and condition of the patient to be treated, the mode of administration used, pathology of the tumour and the required clinical regimen. As a guide, the amount of compound administered by injection is suitably from 0.01 mg/kg to 30 mg/kg, preferably from 0.1 mg/kg to 10 mg/kg.

The routes of administration and dosages described are intended only as a guide since a skilled practitioner will be able to determine readily the optimum route of administration and dosage for any particular patient and condition.

Compounds to be administered may include polypeptides or nucleic acids. The nucleic acids may encode polypeptides or they may encode antisense constructs that inhibit expression of a cellular gene. Nucleic acids may be administered by, for example, lipofection or by viral vectors. For example, the nucleic acid may form part of a viral vector such as an adenovirus. When viral vectors are used, in general the dose administered is between 10⁴ and 10¹⁴ pfu/ml, preferably 10⁶ to 10¹⁰ pfu/ml. The term pfu (“plaque forming unit”) corresponds to the infectivity of a virus solution and is determined by infecting an appropriate cell culture and measuring, generally after 48 hours, the number of plaques of infected cells. The techniques for determining the pfu titre of a viral solution are well documented in the literature.

Any cancer types may be treated by these methods, for example leukaemias, and solid tumours such as breat, ovary, lung, colon, pancreas, testes, liver, brain, muscle and bone tumour. Preferably, the tumour has normal ATR function.

DESCRIPTION OF THE DRAWINGS

FIG. 1

The relationship between ATR, rad3, mei-41, MEC1, TEL1 and ATM

A. Overall structures of ATR, Rad3, Mei-41, Mec1p, Tel1p and ATM.

Legend: open square—Rad3 domain; hatched boxes—kinase domain

B. Dendrogram based on sequence alignments generated by the Clustal method (PAM250) using DNAstar software, rad3/ESR1/mei-41/ATR are more closely related to each other than to ATM and TEL1. Sequences of rad3 and ATM are available in the EMBL database.

The following examples illustrate the invention.

EXAMPLE 1

The rad3 gene of S. pombe is one of six genes absolutely required for the DNA structure checkpoints in S. pombe (Al-Khodairy and Carr, 1992; Al-Khodairy et a. 1994). A sequence refining part of the rad3 gene was reported by Seaton et al. (1992). In attempting to clarify the intron/exon structure of this gene we identified sequencing anomalies at both the 5′ and 3′ ends. We have sequenced the complete gene (see Experimental Procedures) and find that rad3 is capable of encoding a product of 2386 amino acids. The C-terminal region contains the consensus sequences typical of a sub-class of kinases known as lipid kinases, the founder member of which is the p110 catalytic subunit of PI3 kinase (Hiles et al. 1992).

A truncated rad3 clone lacking the amino terminus and the kinase region has been reported to complement the rad3::pR3H1.0 gene disruption mutant of rad3 (Jimenez et al. 1992). This disruption mutant does not remove the potential kinase domain. To clarify the role of this domain, we have created a null mutant by gene replacement. This mutant has amino acids 1477-2271 of rad3, including the kinase consensus domain, replaced by ura4⁺. This strain rad3.d, has identical checkpoint defects and radiation/hydroxyurea sensitivities to the rad3.136 mutant (Nasim and Smith. 1975) and the original rad3::pR3H1.0 disruption mutant (Jimenez et at. 1992: Seaton et al. 1992) (data not shown). We have created three separate point mutants in the putative kinase domain of rad3 and used these in gene replacement experiments to construct strains with defined kinase null mutations. All three stains, rad3.D2230A. rad3.N2235K and rad3.D2249E have phenotypes identical to the rad3.d null mutant da not shown), suggesting that the kinase activity is required for Rad3 function. In the light of our findings, one interpretation of the results of Seaton et al. (1992) and Jimenez et al. (1992) is that the partial clone may show intragenic complementation between the plasmid borne truncated gene and a genomic partial deletion which retains kinase function. Such an interpretation would be consistent with Rad3 acting as a dimer or multimer.

When the kinase Dull allele rad3.D2249E was moderately over-expressed in wild type veils under control of a modified nmt1 promoter (Maundrell, 1990), it caused extreme radiation sensitivity, assayed by UV strip tests, and acted as a dominant negative mutant (data not shown). When the same kinase null construct was expressed at a higher level, it inhibited growth (data not shown). Examination of the cells indicates that division continued very slowly, and at a smaller cell size wild type cells and cells containing empty vector divide at approximately 15 microns, while rad3 and rad3.D2249E over-expressing cells divide at approximately 11.2 microns (data not shown). In S. pombe, this usually indicates an advancement of mitosis.

The Human rad3 Homolog, ATR

To identify a human form of rad3, a combination of methods was applied. Through these approaches, we have cloned the entire coding region of a human gene (see materials and methods). which we have named ATR (ataxia and lad related). ATR is capable of encoding a 2644 amino acid protein which is much more closely related to the products of S. pombe rad3, S. cerevisiae ESR1 (Kato and Ogawa, 1994) and D. melanogaster mei-41 genes (Hari et al. 1995) than to the human ATM and S. cerevisiae Tell proteins (Savitsky et al. 1995; Greenwell et al. 1995) and is likely to be the true homolog of rad3. ESR1 is allelic to the mecI/sad3 checkpoint mutants (Allen et al. 1994; Weinert et al. 1994) which have an equivalent phenotype to rad3. ATR is less closely related to the human checkpoint gene ATM, containing C-terminal putative lipid kinase domain and having a similar overall structure. Sequence alignments demonstrate clearly that the rad3/ESR1(MEC1/SAD3)/mei-41/ATR genes are more closely related to each other than any are to ATR or TEL1, and that ATM is more homologous to TEL1 (FIG. 1).

The ATM gene is expressed in a wide variety of tissues (Savitsky et al. 1995). In S. cerevisiae, ESR1 shows low level expression in mitotic cells but is rapidly in during meiosis 1 (Kato and Ogawa. 1994). Using Northern blot analysis, we have demonstrated that ATR is also weakly expressed in many tissues but that it is mote highly expressed in testis (data not shown). Given that ATP, Rad3 and Esr1p proteins are more highly related to each other than to ATM, the higher ATR expression in testis is consistent with the observation that Esr1p has a role in meiotic recombination (Kato and Ogawa, 1994). Using FISH and PCR analysis, we have mapped ATR to chromosome 3q22-3q25 (data not shown). This region is not associated with known cancer prone syndromes.

In order to further investigate the possibility that Rad3 acts as a multimer, we have created two separate tagged constructs of full length rad3 in pREP based inducible vectors. In one, Rad3 is translated with two myc epitope tags at the N terminus, while in the other these are substituted for a triple HA epitope tag. When both constructs are expressed together in wild type cells, it is possible to co-precipitate the HA tagged Rad3 with the myc specific antibody, and the myc tagged Rad3 with the HA specific antibody (data not shown). This demonstrates that in vivo, the Rad3 protein is capable of self association and is fully consistent with the complementation data of Jimenez et al. (1992).

Although the ATR gene could not complement the phenotype of the rad3 mutants, we have investigated the ability of ATR to form a protein complex with S. pombe Rad3 by expressing both ATR and myc-tagged S. pombe Rad3 in the same yeast cells. Using an anti-ATR antibody (which does not precipitate S. pombe Rad3, see materials and methods) we are able to co-precipitate the yeast protein. We were also able to precipitate the human ATR protein with myc-specific antibodies that recognise the S. pombe Rad3 (data not shown). These data suggest the human and yeast proteins can form a heteromeric-complex, which supports the contention, based on the sequence similarity, of a close functional relationship between these homologues.

Rad3 Proteins have Associated Kinase Activity

Since mutagensis experiments suggest that the kinase activity of the Rad3 proteins in vivo appears to be essential for their function, we have investigated this activity further. Using S. pombe rad3::ura4 cells expressing HA tagged S. pombe Rad3, we have been able to detect a significant protein se activity which precipitates with HA-specific antibodies only when Rad3 is induced and which is not changed following irradiation (data not shown). This activity, which is specific to Rad3 or co-precipitating kinase, appears to reflect phosphorylation of Rad3 itself, since the major band above 200 kD that is phosphorylated can be detected by Western analysis with anti-HA antibody (data not shown). Attempts to identify convenient in vitro substrates such as myelin basic protein. RP-A and several purified S. pombe checkpoint proteins have so far proved unsuccessful. When the IP in vitro kinase assay is performed with cells over-expressing a “kinase-null” D2249E version of Rad3, the associated kinase activity precipitated by HA-specific antibody is significantly reduced (data not shown). There are several possible explanations for this. The measured kinase activity could reflect Rad3 activity directly. In this case the residual activity seen with the kinase dead Rad3 could reflect the fact that it is not unknown for the equivalent D to E mutation in other protein kinases to produce a biologically inert protein with residual in vitro biochemical activity. Alternatively the kinase activity which phosphorylates Rad3 may be due to associated proteins, and these may interact less effectively with the D2249E mutant protein.

Discussion

The checkpoint pathways controlling cell cycle progression following DNA damage or interference in the individual events which comprise the cycle are of considerable importance in maintaining genetic stability and can be considered as pathways which suppress tumorgenesis. Several tumour suppressor genes are intimately involved in subsets of the checkpoint pathways (reviewed in Hartwell and Kastan, 1994), particularly those affecting the transition from G1 into S phase and commitment to the cell cycle. The convergence of the two yeast model systems for checkpoints clearly indicates that the genes involved in these pathways are conserved Our work extends this conservation to metazoan cells, and clarifies the relationship between rad3, ESR1(MEC1/SAD3). mei-41 and the ATM gene.

In this work we demonstrate that the correct sequence of the rad3 gene places its product in the family of protein/lipid kinases related to ATM. Over-expression of kinase-defective rad3 mutant in S. pombe causes a dominant negative phenotype, which suggests that Rad3 is acting as a member of a protein complex whose inter is necessary for checkpoint Onion. This is consistent with the observation that rad1, rad9, rad17, rad26 and has1 deletion mutants all have phenotype indistinguishable from rad3.d (Sheldrick and Carr, 1993). Unexpectedly, unlike the remaining checkpoint rad genes, high level over-expression of either wild type or mutant rad alleles inhibits cell growth and causes mitosis to occur at a reduced cell size, indicative of premature entry into mitosis. This “semi wee” phenotype is not observed in the null mutant, and may indicate interference in a second pathway whose function overlaps with that of Rad3 and acts to inhibit mitosis. A candidate for such a pathway is the ATM/TEL1 pathway which has been shown to have some overlapping functions with the ESR1(MEC1/SAD1) pathway (Morrow et al. 1995).

The structure of ATM is most closely related to the Te11p, which is involved in maintaining telomere length (Greenwell et al., 1995). However, ATM function also appears related to that of the Rad3/Esr1p/mei-41 products. Following the initial discovery of the ATM gene and its sequence relationship to the rad3/ESR1 genes and to TEL1, it was not clear whether, as in many cases in yeast, the gene had duplicated and diverged in yeast, or whether the two yeast proteins defined conserved sub-families of closely related genes. The significant finding of this work is the identification of a human gene, ATR, which is more closely related to rad3/ESR1/mei-41. his defines two sully distinct checkpoint related subfamilies of protein/lipid kinases that are conserved throughout eukaryotic evolution. Although the proteins in these two subfamilies may have some overlapping functions, they probably control different processes. For example: the rad3 sub-family in yeast control all the G1 and G2 DNA damage checkpoints in response to both uv and ionising radiation, and the S phase checkpoint which prevents mitosis following inhibition of replication (Al-Khodairy and Carr, 1992: Allen et al., 1994; Weinert et al., 1994). In contrast, A-T cells have abnormal responses to a narrow range of DNA damaging agents including ionising radiation, biomycin and neocarzinostatin, which produce strand breaks in DNA as a consequence of radical attack. The response to uv and most chemical carcinogens is normal, as is the response to the inhibition of DNA synthesis. It is possible that some or all of the remaining DNA damage checkpoints and the S phase checkpoint are controlled by ATR.

Experimental Procedures

Strains, Plasmids and Media

Standard genetic techniques, growth conditions and media for S. pombe are described in Gutz et al. (1974). S. pombe strain sp011 (ura4.D18, leu.1.32 ade6.704 h⁻) has been described previously (Murray et al. 1992). Plasmid pSUB41 was a gift from S. Subramani (Seaton et al. 1992).

Cloning of S. pombe rad3

A 4.0 kb Kpn1 fragment was excised from pSUB41 and sequenced in both directions to obtain the 5′ rad3 sequence. The 3′ clone was identified from a genomic library (Barbet et al. 1992) by colony hybridisation using a 1 kb 3′ probe derived from the published rad3 sequence, and sequenced in both directions. In this way, the sequence of the entire rad3 gene was assembled.

Null and “Kinase Dead” rad3 Mutants

A construct of rad3, in which the 794 amino acids between aa 1477 and aa 2271 (including kinase domain) were replaced with a ura4+ gene, was created using the methodology describerd in Barbet et at. (1992). A linear fragment of this was used to transform sp011 to uracil prototropy and single copy integration at the rad3 locus was checked by Southern blotting. To create the site specific kinase null mutations, a C-terminsl 3.01 kb bamHI-salI fragment of rad3 was mutated with either (A: GTTTTCGCCATGGCGCGCTCCCAAACCCAA (SEQ ID NO: 5), B: TTCATCAAACAATATCTTTTCGCCATGGCG (SEQ ID NO: 6), or C: CAAAAAGACAGTTGAATTCGACATGGATAG (SEQ ID NO: 7)) in order to introduce either the D2230A, N2235K or D2249E mutations into the kinase domain. Analgous changes have previously been used in the analysis of P13 kinase vps34 of S. cerevisiae (Schu et al. 1993). These fragments were then used to transform the rad3.d null mutant and gene replacements selected by their ability to grow on FOA containing media (Grimm et al. 1988). All strains were checked by Southern blotting. Full length expression constructs of rad3.D2230A were created in pREP1 and pREP41 (Maundrell, 1990) by standard subcloning following introduction of an NdeI site at the ATG and deletion of three internal NdeI sites.

UV Radiation Sensitivity Strip Tests

Expression from REP1 (high) and REP41 (intermediate) was induced by the absence of thiamine for 18 hours prior to plating. Plates were irradiated with a gradient of uv doses down the plate from 0 to 300 Jm⁻² according to the settings on a Stratagene Stratalinker.

Cloning and Expression of ATR

To isolate an appropriate probe for identifying cDNAs corresponding to a human rad3 homologue, degenerate oligonucleotides were designed against the amino acids LGLGDRH (5′ oligo; oDH18)(SEQ ID NO: 13) and HVDF[D/N]C (3′ oligo; oDH-16)(SEQ ID NO: 14) or Rad3/Esr1p. Inosine was incorporated at positions of four-fold degeneracy, and primers were tailed with BamHI (oDH18) and EcoRI (oDH16) to facilitate cloning. DNA sequencen analysis of the ˜100 bp PCR product obtained from amplification of peripheral blood leukocyte cDNA demonstrated significant similarity to MEC1/rad3. This sequence was used to synthesize a non-degenerate primer (oDH-23; GACGCAGAATTCACCAGTCAAAGAATCAAAGAG (SEQ ID NO: 8)) for PCR with an additional degenerate primer (oDH17) designed against the amino acid sequence KFPP[I/V][L/F]Y[Q/E]WF (SEQ ID NO: 12) of Rad3/Esr1p. The 174 bp product of this reaction was used directly to screen a macrophage cDNA library. Four positive clones were isolated (the largest approximately 3 kb.

In parallel, database searches with full length S. pombe rad3 derived from the EMBL database a human cDNA clone, HSAAADPDG, as a potential homologue of rad3, if a single frameshift was allowed for in the 233 bp sequence. This 233 bp sequence is contained within a 1.6 kb clone obtained from Dr. N. Affara, Human Molecular Genetics Research Group, Cambridge University, UK. The entire clone (1.6 kb) was sequenced and lies within the cDNA clones identified by degenerate PCR and library screens. To identify the whole gene, RACE PCR experiments were performed on cDNA derived from placental and thymus mRNA using the instructions provided with a Clontech Marathon Kit. Gene specific primers were derived from the cDNA clones. From these experiments, a 8239 bp cDNA sequence was assembled with an internal ORF of 2644 amino acids, a 79 bp 5′ noncoding region, a 194 bp 3′ noncoding region and a poly A⁻ tail. Parts of the sequence were determined solely by PCR. To avoid errors, clones from a minimum of 3 independent PCR reactions were sequenced in both directions.

The 233 bp sequence corresponds to the sequence of nucleotides 6809 to 7042 (234 nt in total) of Seq. ID No. 1 except for a single base deletion at position 6942. This sequence encodes amino acids 2244 to 2320 of Seq. ID No. 2.

The sequence of the “1.6 kb” insert corresponds to nucleotides 5725 to 7104 (1353 nt) of Seq. ID No. 1, and encodes amino acids 1892 to 2340 of Seq. ID No. 2.

Northern blot hybridisation : a 1.3 kb PCR product was amplified in the presence of ₃₂P-dCTP using primers 279-3 (TGGATGATGA CAGCTGTGTC (SEQ ID NO:9)) and 279-6 (TGTAGTCGCT GCTCAATGTC (SEQ ID NO:10)). A nylon mambrane containing 2 μg of size-fractionated poiy A+ RNA from a variety of human tissue sources (Clontech Laboratories) was probed as recommended by the manufacturer except that the final wash was performed at 55° C. rather than 50° C. to minimize the possibility of cross-hybridization to related sequences.

Mapping ATR

We mapped the ATR gene to chromosome 3 by a combination of fluorescent in situ hybridisation and polymerase chain reaction (PCR) based assays. FISH analysis using a cDNA clone identified the ATR gene on chromosome 3. Two primers (oATR23: GACGCAGAATTCACCAGTCAAAGAATCAAAGAG (SEQ ID NO: 8) and oATR26: TGGTTTCTGAGAACATTCCCTGA (SEQ ID NO:11)) which amplify a 257 bp fragment of the ATR gene were used on DNA derived from humanlrodent somatic cell hybrids containing various human chromosome panels available from the NIGMS Human Genetic Mutant Cell Repository (Drwinga et al. 1993). PCR with the same primers was used to sub-localise ATR to a specific region on chromosome 3. The templates for these amplifications consisted of DNA samples from patients with truncations along chromosome 3 (Leach et al. 1994).

Immunoprecipitation (IP) and Kinase Assays with Rad3

The S. pombe rad3 and human ATR genes were cloned into pREP41 expression vector for complementation studies. To -tag the proteins, versions of these vectors containing in-frame N terminal tag sequences, either a double myc or a triple HA tag, were used (Griffiths et al. 1995). Tagged proteins were expressed by growing in media without thiamine (Maundrell. 1990). Yeast cells lysed in lysis buffer (25 mM Tris.Cl pH 7.5. 60 mM B-glycerophosphate, 0.1 mM Na₃VO₄, 1% Triton X-100, 50 mM MaCl, 2 mM EDTA. 50 mM NaF, 1 mM phenylmethylsulfonyl fluoride [PMSF], 5 μg/ml leupeptin, 5 μg/ml aprotinin, 1 mM DTT) by the addition of glass beads followed by treatment in a dismembrinator for 2 minutes. For IP's 300 μg total protein extract was incubated on ice with the appropriate antibody for 30 min and the immune complexes precipitated by mixing with Protein G beads for a further 30 min at 4° C. For kinase assays, the immune complexes were washed 4 timed with Lysis buffer, once with Kinase Buffer (25 mM Hepes pH7.7; 50 mM KCl; 10 mM MgCl₂; 0.1% NP-40; 2% glycerol; 1 mM DTT), and incubated in Kinase Buffer with 10 μM ATP [50 Ci/mmol]) for 15 minutes at 30° C. The reactions were stopped with 20 ul 2×SDS sample buffer prior to separation on 6% polyacrylamide gels. Rad3 IP's contained several phosphorylated products, including one which comigrated with Rad3 protein itself on Western analysis.

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Sequence Information Sequence ID No, 1: ATR seq    1 GCGCTCTTCCGGCAGCGGTACGTTTGGAGACGCCGGGAACCCGCGTTGGCGTGGTTGACTACTGCCTCGCAGCCT   75   76 CAGCATGGGGGAACATGGCCTGGAGCTGGCTTCCATGATCCCCGCCCTGCGGGAGCTGGGCAGTGCCACACCAGA  150  151 GGAATATAATACAGTTGTACAGAAGCCAAGACAAATTCTGTGTCAATTCATTGACCGGATACTTACAGATGTAAA  225  226 TGTTGTTGCTGTAGAACTTGTAAAGAAAACTGACTCTCAGCCAACCTCCGTGATGTTGCTTGATTTCATCCAGCA  300  301 TATCATGAAATCCTCCCCACTTATGTTTGTAAATGTGAGTGGAAGCCATGAGCGCAAAGGCAGTTGTATTGAATT  375  376 CAGTAATTGGATCATAACGAGACTTCTGCGGATTGCAGCAACTCCCTCCTGTCATTTGTTACACAAGAAAATCTG  450  451 TGAAGTCATCTGTTCATTATTATTTCTTTTTAAAAGCAAGAGTCCTGCTATTTTTGGGGTACTCACAAAAGAATT  525  525 ATTACAACTTTTTGAAGACTTGGTTTACCTCCATAGAAGAAATGTGATGGGTCATGCTGTGGAATGGCCAGTGGT  600  601 CATGAGCCGATTTTTAAGTCAATTAGATGAACACATGGGATATTTACAATCAGCTCCTTTGCAGTTGATGAGTAT  675  676 GCAAAATTTAGAATTTATTGAAGTCACTTTATTAATGGTTCTTACTCGTATTATTGCAATTGTGTTTTTTAGAAG  750  751 GCAAGAACTCTTACTTTGGCAGATAGGTTGTGTTCTGCTAGAGTATGGTAGTCCAAAAATTAAATCCCTAGCAAT  825  826 TAGCTTTTTAACAGAACTTTTTCAGCTTGGAGGACTACCAGCACAACCAGCTAGCACTTTTTTCAGCTCATTTTT  900  901 GGAATTATTAAAACACCTTGTAGAAATGGATACTGACCAATTGAAACTCTATGAAGAGCCATTATCAAAGCTGAT  975  976 AAAGACACTATTTCCCTTTGAAGCAGAAGCTTATAGAAATATTGAACCTGTCTATTTAAATATGCTGCTGGAAAA 1050 1051 ACTGTCTGTCATGTTTGAAGACGGTGTGCTCATGCGGCTTAAGTCTGATTTGCTAAAAGCAGCTTTGTGCCATTT 1125 1126 ACTGCAGTATTTCCTTAAATTTGTGCCAGCTGGGTATGAATCTGCTTTACAAGTCAGGAAGGTCTATGTGAGAAA 1200 1201 TATTTGTAAAGCTCTTTTGGATGTGCTTGGAATTGAGGTAGATGCAGAGTACTTGTTGGGCCCACTTTATGCAGC 1275 1276 TTTGAAAATGGAAAGTATGGAAATCATTGAGGAGATTCAATGCCAAACTCAACAGGAAAACCTCAGCAGTAATAG 1350 1351 TGATGGAATATCACCCAAAAGGCGTCGTCTCAGCTCGTCTCTAAACCCTTCTAAAAGAGCACCAAAACAGACTGA 1425 1426 GGAAATTAAACATGTGGACATGAACCAAAAGAGCATATTATGGAGTGCACTGAAACAGAAAGCTGAATCCCTTCA 1500 1501 GATTTCCCTTGAATACAGTGGCCTAAAGAATCCTGTTATTGAGATGTTAGAAGGAATTGCTGTTGTCTTACAACT 1575 1576 GACTGCTCTGTGTACTGTTCATTGTTCTCATCAAAACATGAACTGCCGTACTTTCAAGGACTGTCAACATAAATC 1650 1651 GAAGAAGAAACCTTCTGTAGTGATAACTTGGATGTCATTGGATTTTTACACAAAAGTGCTTAAGAGCTGTAGAAG 1725 1726 TTTGTTAGAATCTGTTCAGAAACTGGACCTGGAGGCAACCATTGATAAGGTGGTGAAAATTTATGATGCTTTGAT 1800 1801 TTATATGCAAGTAAAGACTTCATTTGAAGATCATATCCTGGAAGATTTATGTGGTATGCTCTCACTTCCATGGAT 1875 1876 TTATTCCCATTCTGATGATGGCTGTTTAAAGTTGACCACATTTGCCGCTAATCTTCTAACATTAAGCTGTAGGAT 1950 1951 TTCAGATAGCTATTCACCACAGGCACAATCACGATGTGTGTTTCTTCTGACTCTGTTTCCAAGAAGAATATTCCT 2025 2026 TGAGTGGAGAACAGCAGTTTACAACTGGGCCCTGCAGAGCTCCCCTGAAGTAATCCGGGCTAGTTGTGTTAGTGG 2100 2101 ATTTTTTATCTTATTGCAGCAGCAGAATTCTTGTAACAGAGTTCCCAAGATTCTTATAGATAAAGTCAAAGATGA 2175 2176 TTCTGACATTGTCAAGAAAGAATTTGCTTCTATACTTGGTCAACTTGTCTGTACTCTTCACGGCATGTTTTATCT 2250 2251 GACAAGTTGTTTAACAGAACCTTTCTCTGAACACGGACATGTGGACCTCTTCTGTAGGAACTTGAAAGCCACTTC 2325 2326 TCAACATGAATGTTCATCTTCTCAACTAAAAGCTTCTGTCTGCAAGCCATTCCTTTTCCTACTGAAAAAAAAAAT 2400 2401 ACCTAGTCCAGTAAAACTTGCTTTCATAGATAATCTACATCATCTTTGTAAGCATCTTGATTTTAGAGAAGATGA 2475 2476 AACAGATGTAAAAGCAGTTCTTGGAACTTTATTAAATTTAATGGAAGATCCAGACAAAGATGTTAGAGTGGCTTT 2500 2551 TAGTGGAAATATCAAGCACATATTGGAATCCTTGGACTCTGAAGATGGATTTATAAAGGAGCTTTTTGTCTTAAG 2625 2626 AATGAAGGAAGCATATACACATGCCCAAATATCAAGAAATAATGAGCTGAAGGATACCTTGATTCTTACAAGACC 2700 2701 GGATATTGGAAGGGCCGCAAAAGGAGATTTGGTACCATTTGCACTCTTACACTTATTGCATTGTTTGTTATCCAA 2775 2776 GTCAGCATCTGTCTCTGGAGCAGCATACACAGAAATTAGAGCTCTGGTTGCAGCTAAAAGTGTTAAACTGCAAAG 2850 2851 TTTTTTCAGCCAGTATAAGAAACCCATCTGTCAGTTTTTGGTAGAATCCCTTCACTCTAGTCAGATGACAGCACT 2925 2926 TCCGAATACTCCATGCCAGAATGCTGCCGTGCGAAAACAAGATGTGGCTCACCAGAGAGAAATGGCTTTAAATAC 3000 3001 GTTGTCTGAAATTGCCAACGTTTTCGACTTTCCTGATCTTAATCGTTTTCTTACTAGGACATTACAAGTTCTACT 3075 3076 ACCTGATCTTGCTGCCAAAGCAAGCCCTGCAGCTTCTGCTCTCATTCGAACTTTAGGAAAACAATTAAATGTCAA 3150 3151 TCGTAGAGAGATTTTAATAAACAACTTCAAATATATTTTTTCTCATTTGGTCTGTTCTTGTTCCAAAGATGAATT 3225 3226 AGAACGTGCCCTTCATTATCTGAAGAATGAAACAGAAATTGAACTGGGGAGCCTGTTGAGACAAGATTTCCAAGG 3300 3301 ATTGCATAATGAATTATTGCTGCGTATTGGAGAACACTATCAACAGGTTTTTAATGGTTTGTCAATACTTGCCTC 3375 3376 ATTTGCATCCAGTGATGATCCATATCAGGGCCCGAGAGATATCATATCACCTGAACTGATGGCTGATTATTTACA 3450 3451 ACCCAAATTGTTGGGCATTTTGGCTTTTTTTAACATGCAGTTACTGAGCTCTAGTGTTGGCATTGAAGATAAGAA 3525 3526 AATGGCCTTGAACAGTTTGATGTCTTTGATGAAGTTAATGGGACCCAAACATGTCAGTTCTGTGAGGGTGAAGAT 3600 3601 GATGACCACACTGAGAACTGGCCTTCGATTCAAGGATGATTTTCCTGAATTGTGTTGCAGAGCTTGGGACTGCTT 3675 3676 TGTTCGCTGCCTGGATCATGCTTGTCTGGGCTCCCTTCTCAGTCATGTAATAGTAGCTTTGTTACCTCTTATACA 3750 3751 CATCCAGCCTAAAGAAACTGCAGCTATCTTCCACTACCTCATAATTGAAAACAGGGATGCTGTGCAAGATTTTCT 3825 3826 TCATGAAATATATTTTTTACCTGATCATCCAGAATTAAAAAAGATAAAAGCCGTTCTCCAGGAATACAGAAAGGA 3900 3901 GACCTCTGAGAGCACTGATCTTCAGACAACTCTTCAGCTCTCTATGAAGGCCATTCAACATGAAAATATCGATCT 3975 3976 TCGTATTCATGCTCTTACAAGCTTGAAGGAAACCTTGTATAAAAATCAGGAAAAACTGATAAAGTATGCAACAGA 4050 4051 CAGTGAAACAGTAGAACCTATTATCTCACAGTTGGTGACAGTGCTTTTGAAAGGTTGCCAAGATGCAAACTCTCA 4125 4126 AGCTCGGTTGCTCTGTGGGGAATGTTTAGGGGAATTGGGGGCGATAGATCCAGGTCGATTAGATTTCTCAACAAC 4200 4201 TGAAACTCAAGGAAAAGATTTTACATTTGTGACTGGAGTAGAAGATTCAAGCTTTGCCTATGGATTATTGATGGA 4275 4276 GCTAACAAGAGCTTACCTTGCGTATGCTGATAATAGCCGAGCTCAAGATTCAGCTGCCTATGCCATTCAGGAGTT 4350 4351 GCTTTCTATTTATGACTGTAGAGAGATGGAGACCAACGGCCCAGGTCACCAATTGTGGAGGAGATTTCCTGAGCA 4425 4426 TGTTCGGGAAATACTAGAACCTCATCTAAATACCAGATACAAGAGTTCTCAGAAGTCAACCGATTGGTCTGGAGT 4500 4501 AAAGAAGCCAATTTACTTAAGTAAATTGGGTAGTAACTTTGCAGAATGGTCAGCATCTTGGGCAGGTTATCTTAT 4575 4576 GACCATCTATCTTCTTCCACATATTCTGGTGTATGTCTTACTGGGTTGTAATCAAGAAGATCAGCAGGAGGTTTA 4725 4725 TGCAGAAATTATGGCAGTTCTAAAGCATGACGATCAGCATACCATAAATACCCAAGACATTGCATCTGATCTGTG 4800 4801 TCAACTCAGTACACAGACTGTGTTCTCCATGCTTGACCATCTCACACAGTGGGCAAGGCACAAATTTCAGGCACT 4875 4876 GAAAGCTGAGAAATGTCCACACAGCAAATCAAACAGAAATAAGGTAGACTCAATGGTATCTACTGTGGATTATGA 4950 4951 AGACTATCAGAGTGTAACCCGTTTTCTAGACCTCATACCCCAGGATACTCTGGCAGTAGCTTCCTTTCGCTCCAA 5025 5026 AGCATACACACGAGCTGTAATGCACTTTGAATCATTTATTACAGAAAAGAAGCAAAATATTCAGGAACATCTTGG 5100 5101 ATTTTTACAGAAATTGTATGCTGCTATGCATGAACCTGATGGAGTGGCCGGAGTCAGTGCAATTAGAAAGGCAGA 5175 5176 ACCATCTCTAAAAGAACAGATCCTTGAACATGAAAGCCTTGGCTTGCTGAGGGATGCCACTGCTTGTTATGACAG 5250 5251 GGCTATTCAGCTAGAACCAGACCAGATCATTCATTATCATGGTGTAGTAAAGTCCATGTTAGGTCTTGGTCAGCT 5325 5326 GTCTACTGTTATCACTCAGGTGAATGGAGTGCATGCTAACAGGTCCGAGTGGACAGATGAATTAAACACGTACAG 5400 5401 AGTGGAAGCAGCTTGGAAATTGTCACAGTGGGATTTGGTGGAAAACTATTTGGCAGCAGATGGAAAATCTACAAC 5475 5476 ATGGAGTGTCAGACTGGGACAGCTATTATTATCAGCCAAAAAAAGAGATATCACAGCTTTTTATGACTCACTGAA 5550 5551 ACTAGTGAGAGCAGAACAAATTGTACCTCTTTCAGCTGCAAGCTTTGAAAGAGGCTCCTACCAACGAGGATATGA 5625 5626 ATATATTGTGAGATTGCACATGTTATGTGAGTTGGAGCATAGCATCAAACCACTTTTCCAGCATTCTCCAGGTGA 5700 5701 CAGTTCTCAAGAAGATTCTCTAAAGTGGGTAGCTCGACTAGAAATGACCCAGAATTCCTACAGAGCCAAGGAGCC 5775 5776 TATCCTGGCTCTCCGGAGGGCTTTACTAAGCCTCAACAAAAGACCAGATTACAATGAAATGGTTGGAGAATGCTG 5850 5851 GCTGCAGAGTGCCAGGGTAGCTAGAAAGGCTGGTCACCACCAGACAGCCTACAATGCTCTCCTTAATGCAGGGGA 5925 5926 ATCACGACTCGCTGAACTGTACGTGGAAAGGGCAAAGTGGCTCTGGTCCAAGGGTGATGTTCACCAGGCAATAAT 6000 6001 TGTTCTTCAAAAAGGTGTTGAATTATGTTTTCCTGAAAATGAAACCCCACCTGAGGGTAAGAACATGTTAATCCA 6075 6076 TGGTCGAGCTATGCTACTAGTGGGCCGATTTATGGAAGAAACAGCTAACTTTGAAAGCAATGCAATTATGAAAAA 6150 6151 ATATAAGGATGTGACCGCGTGCCTGCCAGAATGGGAGGATGGGCATTTTTACCTTGCCAAGTACTATGACAAATT 6225 6226 GATGCCCATGGTCACAGACAACAAAATGGAAAAGCAAGGTGATCTCATCCGGTATATAGTTCTTCATTTTGGCAG 6300 6301 ATCTCTACAATATGGAAATCAGTTCATATATCAGTCAATGCCACGAATGTTAACTCTATGGCTTCATTATGGTAC 6375 6376 AAAGGCATATGAATGGGAAAAAGCTGGCCGCTCCGATCGTGTACAAATGAGGAATGATTTGGGTAAAATAAACAA 6450 6451 GGTTATCACAGAGCATACAAACTATTTAGCTCCATATCAATTTTTGACTGCTTTTTCACAATTGATCTCTCGAAT 6525 6526 TTGTCATTCTCACGATGAAGTTTTTGTTGTCTTGATGGAAATAATAGCCAAAGTATTTCTAGCCTATCCTCAACA 6600 6601 AGCAATGTGGATGATGACAGCTGTGTCAAAGTCATCTTATCCCATGCGTGTGAACAGATGCAAGGAAATCCTCAA 6675 6676 TAAAGCTATTCATATGAAAAAATCCTTAGAGAAGTTTGTTGGAGATGCAACTCGCCTAACAGATAAGCTTCTAGA 6750 6751 ATTGTGCAATAAACCGGTTGATGGAAGTAGTTCCACATTAAGCATGAGCACTCATTTTAAAATGCTTAAAAAGCT 6825 6826 GGTAGAAGAAGCAACATTTAGTGAAATCCTCATTCCTCTACAATCAGTCATGATACCTACACTTCCATCAATTCT 6900 6901 GGGTACCCATGCTAACCATGCTAGCCATGAACCATTTCCTGGACATTGGGCCTATATTGCAGGGTTTGATGATAT 6975 6976 GGTGGAAATTCTTGCTTCTCGGCAGAAACCAAAGAAGATTTCTTTAAAAGGCTCAGATGGAAAGTTCTACATCAT 7050 7051 AGACCGTCATGGTGAAAATATTCTCTTTGATTCTTTGACTGGTGAATGCGTACATGTAGATTTCAATTGTCTTTT 7125 7126 AAGAAAAGATGCAGAGTCTCGTAGAAGAGAACTTCATATTCGAACATATGCAGTTATTCCACTAAATGATGAATG 7200 7201 TGGGATTATTGAATGGGTGAACAACACTGCTGGTTTGAGACCTATTCTGACCAAACTATATAAAGAAAAGGGAGT 7275 7276 GTATATGACAGGAAAAGAACTTCGCCAGTGTATGCTACCAAAGTCAGCAGCTTTATCTGAAAAACTCAAAGTATT 7350 7351 CCGAGAATTTCTCCTGCCCAGGCATCCTCCTATTTTTCATGAGTGGTTTCTGAGAACATTCCCTGATCCTACATC 7425 7426 ATGGTACAGTAGTAGATCAGCTTACTGCCGTTCCACTGCAGTAATGTCAATGGTTGGTTATATTCTGGGGCTTGG 7500 7501 AGACCGTCATGGTGAAAATATTCTCTTTGATTCTTTGACTGGTGAATGCGTACATGTAGATTTCAATTGTCTTTT 7575 7576 CAATAAGGGAGAAACCTTTGAAGTTCCAGAAATTGTGCCATTTCGCCTGACTCATAATATGGTTAATGGAATGGG 7650 7651 TCCTATGGGAACAGAGGGTCTTTTTCGAAGAGCATGTGAAGTTACAATGAGGCTGATGCGTGATCAGCGAGAGCC 7725 7726 TTTAATGAGTGTCTTAAAGACTTTTCTACATGATCCTCTTGTGGAATGGAGTAAACCAGTGAAAGGGCATTCCAA 7800 7801 AGCGCCACTGAATGAAACTGGAGAAGTTGTCAATGAAAAGGCCAAGACCCATGTTCTTGACATTGAGCAGCGACT 7875 7876 ACAAGGTGTAATCAAGACTCGAAATAGAGTGACAGGACTGCCGTTATCTATTGAAGGACATGTGCATTACCTTAT 7950 7951 ACAAGAAGCTACTGATGAAAACTTACTATGCCAGATGTATCTTGGTTGGACTCCATATATGTGAAATGAAATTAT 8025 8026 GTAAAAGAATATGTTAATAATCTAAAAGTAATGCATTTGGTATGAATCTGTGGTTGTATCTGTTCAATTCTAAAG 8100 8101 TACAACATAAATTTACGTTCTCAGCAACTGTTATTTCTCTCTGATCATTAATTATATGTAAAATAATATACATTC 8175 8176 AGTTATTAAGAAATAAACTGCTTTCTTAATAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA            8239 Sequence ID No. 2: ATR protein    1 MGEHGLELASMIPALRELGSATPEEYNTVVQKPRQILCQFIDRILTDVNVVAVELVKKTDSQPTSV            66   67 MLLDFIQHIMKSSPLMFVNVSGSHERKGSCIEFSNWIITRLLRIAATPSCHLLHKKICEVICSLLFLFKSKSPAI  141  142 FGVLTKELLQLFEDLVYLHRRNVMGHAVEWPVVMSRFLSQLDEHMGYLQSAPLQLMSMQNLEFIEVTLLMVLTRI  216  217 IAIVFFRRQELLLWQIGCVLLEYGSPKIKSLAISFLTELFQLGGLPAQPASTFFSSFLELLKHLVEMDTDQLKLY  291  292 EEPLSKLIKTLFPFEAEAYRNIEPVYLNMLLEKLCVMFEDGVLMRLKSDLLKAALCHLLQYFLKFVPAGYESALQ  366  367 VRKVYVRNICKALLDVLGIEVDAEYLLGPLYAALKMESMEIIEEIQCQTQQENLSSNSDGISPKRRRKSSSLNPS  441  442 KRAPKQTEEIKHVDMNQKSILWSALKQKAESLQISLEYSGLKNPVIEMLEGIAVVLQLTALCTVHCSHQNMNCRT  516  517 FKDCQHKSKKKPSVVITWMSLDFYTKVLKSCRSLLESVQKLDLEATIDKVVKIYDALIYMQVNSSFEDHILEDLC  591  592 GMLSLPWIYSHSDDGCLKLTTFAANLLTLSCRISDSYSPQAQSRCVFLLTLFPRRIFLEWRTAVYNWALQSSHEV  666  667 IRASCVSGFFILLQQQNSCNRVPKILIDKVKDDSDIVKKEFASILGOLVCTLHGMFYLTSSLTEPFSEHGHVDLF  671  742 CRNLKATSQHECSSSQLKASVCKPFLFLLKKKIPSPVKLAFIDNLHHLCKHKDFREDETDVKAVLGTLLNLMEDP  816  817 DKDVRVAFSGNIKHILESLDSEDGFIKELFVLRMKEAYTHAQISRNNELKDTLILTTGDIGRAAKGDLVPFALLH  891  892 LLHCLLSKSASVSGAAYTEIRALVAAKSVKLQSFFSQYKKPICQFLVESLHSSQMTALPNTPCQNADVRKQDVAH  966  967 QREMALNTLSEIANVFDFPDLNRFLTRTLQVLLPDLAAKASPAASALIRTLGKQLNVNRREILINNFKYIFSHLV 1041 1042 CSCSKDELERALHYLKNETEIELGSLLRQDFQGLHNELLLRIGEHYQQVFNGLSILASFASSDDPYQGPRDIISP 1116 1117 ELMADYLQPKLLGILAFFNMQLLSSSVGIEDKKMALNSLMSLMKLMGPKHVSSVRVKMMTTLRTGLRFKDDFPEL 1191 1192 CCRAWDCFVRCLDHACLGSLLSHVIVALLPLIHIQPKETAAIFHYLIIENRDAVQDFLHEIYFLPDHPELKKIKA 1266 1267 VLQEYRKETSESTDLQTTLQLSMKAIQHENVDVRIHALTSLKETLYKNQEKLIKYATDSETVEPIISQLVTVLLK 1341 1342 GCQDANSQARKKCGECLGELGAIDPGRLDFSTTETQGKDFTFVTGVEDSSFAYGLLMELTRAYLAYADNSRAQDS 1416 1417 AAYAIQELLSIYDCREMETNGPGHQLWRRFPEHVREILEPHLNTRYKSSQKSTDWSGVKKPIYLSKLGSNFAEWS 1491 1492 ASWAGYLITKVRHDLASKIFTCCSIMMKHDFKVTIYLLPHILVYVLLGCNQEDQQEVYAEIMAVLKHQDQHTINT 1566 1567 QDIASDLCQLSTQTVFSMLDHLTQWARHKFQALKAEKCPHSKSNRNKVDSMSTVDYEDYQSVTRFLDLIPPQDTL 1641 1642 AVASFRSKAYTRAVMHFESFITEKKQNIQEHLGFLQKLYAAMHEPDGVAGVSAIRKAEPSLKEQILEHESLGLLR 1716 1717 DATACYDRAIQLEPDQIIHYHGVVKSMLGLGQLSTVITQVNGVHANRSEWTDELNTYRVEAAWKLSQWDLVENYL 1791 1792 AADGKSTTWSVRLGQLLLSAKKRDITAFYDSLKLVRAEQIVPLSAASFERGSYQRGYEYIVRLHMLCELEHSIKP 1866 1867 LFQHSPGDSSQEDSLNWVARLEMTQNSYRAKEPILALRRALLSLNKRPDYNEMVGECWLQSARVARKAGHHQTAY 1941 1942 NALLNAGESRLAELYVERAKWLWSKGDVHQALIVLQKGVELCFPENETPPEGKNMLIHGRAMLLVGRFMEETANF 2016 2017 ESNAIMKKYKDVTACLPEWEDGHFYLAKYYDKLMPMVTDNKMEKQGDLIRYIVLHFGRSLQYGNQFIYQSMPRML 2091 2092 TLWLDYGTKAYEWEKAGRSDRVQMRNDLGKINKVITEHTNYLAPYQFLTAFSQLISRICHSHDEVFVVLMEIIAK 2166 2167 VFLAYPQQAMWMMTAVSKSSYPMRVNRCKEILNKAIHNKKSLEKFVGDATRLTDKLLELCNKPVDGSSSTLSMST 2241 2242 HFKMLKKLVEEATFSEILIPLQSVMIPTLPSILGTHANHASHEPFPGHWAYIAGFDDMVEILASLQKPKKISLKG 2316 2317 SDGKFYIMMCKPKDDLRKDCRLMEFNSLINKCLRKDAESRRRELHIRTYAVIPLNDECGIIEWVNNTAGLRPILT 2391 2392 KLYKEKGVYMTGKELRQCMLPKSAALSEKLKVFREFLLPRHPPIFHEWFLRTFPDPTSWYSSRSAYCRSTAVMSM 2466 2467 VGYILGLGDRHGENILFDSLTGECVHVDFNCLFNKGETFEVPEIVPFRLTHNMVNGMGPMGTEGLFRRACEVTNR 2541 2542 LMRDQREPLMSVLKTFLHDPLVEWSKPVKGHSKAPLMETGEVVNEKAKTHVLDIEQRLQGVIKTRNRVTGLPLSI 2616 2617 EGHVHYLIQEATDENLLCQMYLGWTPYM                                                2664 Sequence ID No. 3: rad3.seq    1 GGTACCAAGTAAAAACTGCTTAGTAAGTATAAAACACAGAAGAATCCGCGATCTAGTGAACCAATGCCCTGCGTA   75   76 TGACGCTCCACTGACGCTATAGTCAATGAGAACTAGGATGTGCGATTATAACTTATCTTTTCAATATTTTCTTAT  150  151 TATTTATTTAAGAAATAATTGAATTAAAACTCATTTCTTCTTTTATTAGCCGTAAAATAGCTTATTTTCTCTCCT  225  226 ACTACCTTTCAACAATAACTTTTTTTTTTGTTTATTGACCATTATAATCACATCAAAAGTCAAAAAATTCAATCA  300  301 TTATCAGAAACATCCAGCCTAATATTACTTAAAAGTTAGTTTCCTCTGAAAATTCAGTATCACAAAAGCTCGTTA  375  376 ATTAGCATCGCTCGATACTTAGTGCACCATGCATCTTCCTTTACCTCGTGAGTGGAAATCGATTTGATAATCGAT  450  451 TGCCACTTTTCGCATAATTCTATTGAGATATTTTATTACTTACAATCGTCTTTTATAAATGCTCAAGACTTTGAA  525  526 CGCGCGTGTTGCGTTTTAAAAAGGCCTTTTTTTGAATTGAATCAATGGTTTGATATAGTATGAGCCAACACGCAA  600  601 AAAGGAAAGCTGGGTCACTCGATCTTTCACCCAGAGGCTTAGATGACAGACAGGCTTTCGGACAGCTTTTGAAAG  675  676 AAGTATTAGCATTAGACAAAGAACATGAGTTAGGTAGAAGTAATTCTTTACCATCTATGACCTCCGAGCTTGTTG  750  751 AAGTTTTAATTGAAGTTGGTCTTCTAGCTTTTAAACATGATGATTCAAAATCTGAATTTATCTCTCCTAAGATGC  825  826 TAAAAGAAGCCCATCTCTCTCTACAAGCGTTAATGCTAATCTTAAAAAGGTCTCCGACAGTTTTGCGGGAGATTA  900  901 AATCATCTGTTACTCTTTTGGATTGGATTTTACCCAGGACTATATCATTGTTTGCTGATATTCGTTTTATTAAGT  975  976 TATTTGACTCATTAAAAGAGTTTCATAAGCTAATTTATCAGCTAATCAGTGAAAAGTCATTCCTATGGGACTTAT 1050 1051 ATGCTTCGTTTATGCGTTATTGGAAATATTATATTACAAACGTTTCTTCTATAGTTCTCCAAATCACTAATGCTA 1125 1126 CATTCCCTTACAAGATGCCCTCACCCAATTCTCAACCATTGCAGAGTATCTCCCCAAATTATCCAACCCATCGAG 1200 1201 AGGACAAATTTGATTTACTTATCATTAATATAGAGGAGGCTTGTACATTTTTCTTTGAAAGTGCCCATTTTTTTG 1275 1276 CACAATGCTCATATTTAAAGAAATCCAATTTTCCTAGTCCACCTCTCTTTACAGCGTGGACTTGGATCAAGCCAT 1350 1351 GTTTTTTTAATTTTGTTATTTTATTAAAACGAATCAGCATCGGAGACTCACAGCTCTTTCTACATTTGCATTCAC 1425 1426 GTATAGTCCAAACTTTATGCTGTTTTTCCTTGAATTTTATATATCATGGCCTTCCCATTTGTGAAAAATCTAAAC 1500 1501 ATATTTTAATGTCCTCCATCAACTTAACATTGGGATCATTGAAGAAAACTTATACAGTTGCTAATACTGCTATAT 1575 1576 CTCTTTTTTTTCTCTCTTTATTTGTTTTACCCAAAACTGTAGCTGGTCTATTCTATCCTTTTGGGGTTTCCTTAC 1650 1651 TTTCTGACTTCAAGGTATTAGAGCAACTTGAACCAGATTCTGATCTCAAAAAGGCAATAATATTATTTAAGTGCA 1725 1726 GATACCAAAGTTCAGAAATAGATCAAACAACTCTCCGTGCTTTTGGCGAAATTTGTACTGGTAAACTTGAAAACA 1800 1801 CGTTGTTTTCTAACTCTGAATTAAACCTTTTTCTTTTACATTATCTTTCCTTGGACAATGACTTGTCAAATATTC 1875 1876 TTAAAGTGGATTTCCAGAATGGTCATAACATATGTACATTTGCAAAATGGTGTATAAACAACAACTTAGATGAAC 1950 1951 CGTCTAATTTAAAGCACTTTCGTGAAATGTTAGATTATTATAGCTCTCATAATGTTACAATAAGTGAGGACGACC 2025 2026 TGAAGAACTTCTCTTTAGTTTTGTGTACTCATGTTGCAAAGGTGAATGAGAAAACAAATAGTATTTTCCGCACAT 2100 2101 ATGAAGTACATGGTTGTGAAGTTTGTAACTCATTTTGTTTACTATTTGATGAGCGGTCGCCTTTTAAAATTCCTT 2175 2176 ATCACGAATTGTTTTGTGCATTGCTAAAAAATCCCGCAATAATTTCCTCTTCTGTTAAACAATCATTGTTGCTTG 2250 2251 ATGGCTTTTTTCGGTGGAGCCAGCATTGCTCAAACTTTAATAAAGAATCAATGTTAAGTTTAAGAGAATTTATTA 2325 2326 TGAAAGCATTAGCCAGTACTTCAAGATGTTTACGTGTTGTTGCTGCAAAAGTTTTGCCCATTTTCATTAAGGGAC 2400 2401 CTAATAATCTTGATATAGTTGAATTTCACAAGGAAAGTAAAGCCTTGATTTTTAATACGTTGAAAATATTGGCGG 2475 2476 TGGAAAATACAGCTATTTTAGAA A C G GTCAT T C T TTCCTGGATCTCCTTATCTAGAGTGGTAGAAGAAGAAGAAT 2550 2551 TACATTTTGTACTATTGGAAGTTATATCTTCTGTGATAAACAGCGGAATATTTTATCAAGGCATTGGTCTCAGCG 2625 2626 CTCTGCAACAAATTGCCTCGACGCGTCATATATCCGTTTGGCAATTACTTTCTCCATATTGGCCAACAGTGTCCG 2700 2701 TTGCGATTGTCCAAGGTATGGGTAAAAAACCGAACATAGCCAGTTTATTTGCTCAGCTTATGAATATTTCCGAGG 2775 2776 GCGATTTTCTTATTCGAACAGAGGCGTACACTTTACCATTCCTTGTACTTACTAAAAACAAAGCGTTAATAGTAC 2850 2851 GTATAGCTGAACTTTCACAAAGTGATGTTGCTACTTTGTGCCTTACCAATATGCATAAAATCCTTGCTTCGCTAC 2925 2926 TTACTACGGATCATCCTAATTTGGAAGAGAGTGTGATGCTTCTTCTTTCACTGGCCACTTCTGATTTTGAAAAAG 3000 3301 TTGATTTAACGTCTTTGTTACGCTCTGATCCTATTTCTATTACTGTGGAGTTGTTACAGCTTTATCAGAATGATG 3075 3076 TTCCTCATGAAAAAATTGAAAATGCTTTAAGAAAGGTAGCAATGATTCTGTCTCAAGTGGTTAATGACGAAGACT 3150 3151 TGAGCAATAAGGAATTACTTTATGATTTTTTTAATAATCACATTTTGGGTATCTTAGCAGAATTTTCTAATATCC 3225 3226 TTAACGACCTGAAAGGAAAGACTTCAATTAATGAAAAGATTAAGACAATTGTCGGCATTGAAAAAATGTTATCTT 3300 3301 TATGTGGAGGTGCAGTCAAACTTGGATTACCACAGATACTTTCTAATTTACAAAGTGCTTTTCAAAATGAGCACT 3375 3376 TAAGGTTTTATGCAATCAAAGCTTGGTTCAGTTTGATATTAGCAACCAAGGAGCCCGAGTATAGTTCAATTGCTG 3450 3451 GTTTAAGTCTTGTAATTTTACCTCCTTTATTCCCTTATTTAGAACCACAAGAAGCAGAGCTAGTAATTCAAATAT 3525 3526 TTGATTTTATTTCTTCTGACACACACAAGTGCCTACAAGGATTAAAGTGGGCTATCCCCACCAGTCTGGATTCAG 3600 3601 CGTGCTTTAGCCTTAAGGCTAAAGAAATATTCTGTTCGCTTCAAAATGAAGATTTTTACTCTGAGCTTCAAAGTA 3675 3676 TAATTAAGTGTTTAACTAACGAAAATGAGCCAGTTTGTTATTTAGGTTTACAAAAATTAGAACTTTTTTTTCAAG 3750 3751 CCAAGGTGGACGAGTTACATGACACACTAAATTTGGACATATCCAACGAAGTTCTGGACCAATTACTAAGATGCC 3825 3826 TTTTAGATTGTTGTGTAAAATATGCTTCAACAAATATGCAAATATCATATCTTGCTGCAAAAAATCTTGGTGAAT 3900 3901 TGGGTGCGATAGATCCCAGCCGCGCCAAGGCTCAACATATTATTAAAGAAACAGTTGTTCTTGATAACTTTGAAA 3975 3976 ACGGAGAAGAAAGTTTGAAGTTTATTCTAGATTTTATGCAATCGCAGTTAATTCCAGCTTTCCTTGTTACTACTG 4050 4051 ATACTAAAGCACAAGGTTTTCTTGCCTATGCTCTGCAAGAGTTTCTAAAGCTTGGTGGATTCAAGTCCGCAGTGA 4125 4126 TTAATAAAAAAAAGGGACTAACTGTGGTAACAGAACATTGGATGTCTTTGCCTGATTTATCCAAACGTGTGCTTA 4200 4201 TACCATTTTTAACTTCCAAGTATCATTTAACACCAATCCCCAAAATTGACATTCGGTACCCTATTTATAAAGAAA 4275 4276 ATGTTACTATTCATACTTGGATGCAGTTGTTTTCTCTTAAATTGATGGAGTACGCCCATTCGCAAAACGCTGAAA 4350 4351 AAATATTTGGTATTTGTTCGAAAGTAGTGAAAGACCAAGAGGTTAACATTCCCTGTTTTCTTCTTCCCTTTCTTG 4425 4426 TTTTAAATGTTATTTTAACCGAGTCAGAACTGGAAGTTAATAAAGTCATTGAAGAATTCCAGCTTGTTATTAATC 4500 4501 AACCGGGACCTGATGGATTAAATTCCGTGGGGCAACAAAGATACACCTCATTTGTAGATGTATTTTTTAAGATTG 4575 4576 TGGATTACCTTAACAAATGGCTTCGCATGCGAAAGAAGAGGAATTGGGATAGACGTTCTGCCATTGCAAGGAAAG 4650 4651 AGAACCGTTATATGTCGGTGGAAGATGCTACCTCTCGAGAATCATCGATCTCAAAAGTTGAGTCATTTCTTTCTC 4725 4726 GATTTCCTTCAAAAACATTAGGTATTCTCTCTTTAAATTGTGGATTTCATGCTCGTGCATTGTTTTATTGGGAGC 4800 4801 AACACATACGTAATGCTACAGCTCCATATGCAGCTTTAGAGTCCGATTATAGAGTTTTGCAGGAAATATATGCTG 4875 4876 GAATTGATGATCCAGATGAAATCGAAGCAGTGTCTTTAAATTTCCATGATTACTCGTTTGATCAACAACTCCTTT 4950 4951 TACATGAAAATTCAGGAACATGGGACTCGGCTTTGAGTTGTTACGAAATTATTATTCAAAAGGATCCTGAAAATA 5025 5026 AAAAGGCGAAAATCGGTTTGCTTAACAGCATGCTGCAATCGGGGCATTATGAATCTCTTGTTTTGAGTTTAGATT 5100 5101 CTTTTATAATCAATGACAACCACGAGTATTCGAAGATGTTAAATTTGGGTATTGAAGCTTCATGGCGTTCGCTAT 5175 5176 CTATTGATTCGTTAAAAAAGTGTCTTTCAAAAAGCAACTTGGAATCTTTCGAAGCTAAATTGGGTAGCATATTTT 5250 5251 ACCAATACCTACGGAAGGATTCTTTTGCTGAATTGACGGAGCGGCTGCAACCCTTGTACGTTGATGCTGCTACAG 5325 5326 CAATTGCAAACACAGGAGAAAATTCAGCCTATGATTGTTATGATATTTTATCTAAGCTGCACGCAATTAATGACT 5400 5401 TTAGTAGGATTGCTGAAACTGACGGAATTGTTTCCGACAATCTTGATATTGTTCTTCGCCGTCGGCTTAGCCAAG 5475 5476 TAGCTCCGTACGGTAAATTCAAGCACCAAATCCTGTCCACTCACTTAGTTGGCTATGAAAAATTTGAAAACACGA 5550 5551 AGAAAACTGCTGAAATATATCTCGAGATTGCAAGAATATCTCGAAAAAATGGTCAATTTCAAAGAGCCTTCAATG 5625 5626 CCATCCTCAAAGCAATGGATTTAGATAAACCGCTAGCAACAATAGAGCACGCACAATGGTGGTGGCATCAAGGGC 5700 5701 AACATCGTAAAGCTATTTCTGAATTGAATTTTTCGCTTAATAACAACATGTTTGATTTGGTTGATGAGCATGAAG 5775 5776 AAAGACCTAAAAATCGTAAAGAAACTTTAGGAAATCCACTTAAAGGAAAAGTGTTCTTGAAACTTACAAAATGGC 5850 5851 TCGGAAAAGCTGGCCAACTGGGATTGAAGGATTTGGAGACGTATTATCATAAAGCGGTAGAGATTTA CT CAGAAT 5925 5926 GTGAGAATACGCATTATTATCTTGGCCATCATCGAGTTTTAATGTATGAAGAAGAACAAAAGCTCCCAGTTAATG 6000 6001 AACAGAGCGAACGATTTTTAAGTGGTGAGTTAGTAACTCGCATAATTAACGAATTTGGTCGATCTTTGTACTATG 6075 6076 GTACAAATCATATATATGAAAGTATGCCAAAATTGCTCACACTGTGGCTTGATTTTGGGGCCGAAGAACTTCGCT 6150 6151 TATCTAAAGATGACGGCGAAAAGTACTTTCGTGAACACATTATCTCTTCGAGAAAAAAATCTTTGGAACTTATGA 6225 6226 ATTCGAATGTTTGTCGCCTTTCTATGAAAATTCCTCAATACTTTTTTGTCCTTGCATTATCCCAAATGATATCCA 6300 6301 GAGTATGCCATCCAAATAATAAAGTTTATAAAATTTTGGAACATATAATTGCAAACGTTGTAGCATCTTATCCTG 6375 6376 GGGAGACGCTATGGCAATTAATGGCAACAATAAAATCGACTTCTCAAAAGCGCTCGCTTCGTGGAAAAAGCATTT 6450 6451 TAAATGTTTTACATTCTAGGAAGCTTTCTATGTCTTCCAAAGTTGATATAAAAGCACTCAGTCAATCTGCAATTC 6525 6526 TCATTACTGAAAAGTTAATCAATTTGTGCAATACAAGGATTAACAGTAAATCTGTAAAAATGAGCTTAAAGGATC 6600 6601 ATTTTCGGCTTTCTTTTGATGATCCGGTAGATTTAGTCATTCCTGCTAAATCATTTTTAGACATTACTTTACCAG 6675 6676 CTAAAGATGCTAACAGAGCTAGTCATTATCCATTTCCAAAAACTCAGCCTACTCTGTTGAAATTTGAGGATGAGG 6750 6751 TGGATATAATGAACTCTCTTCAAAAACCAAGAAAAGTGTACGTTAGAGGTACGGATGGCAACTTATACCCATTCT 6825 6826 TGTGCAAACCCAAAGATGATCTTCGTAAGGATGCTAGATTGATGGAATTTAATAATCTTATTTGTAAAATATTGA 6900 6901 GGAAAGATCAAGAAGCGAACAGAAGGAACTTGTGTATTAGAACTTATGTTGTTATTCCTTTAAATGAAGAATGCG 6975 6976 GATTTATCGAATGGGTAAATCATACTCGTCCATTTAGAGAAATTTTGTTAAAAAGCTATAGACAGAAAAACATTC 7050 7051 CCATATCATATCAAGAAATCAAAGTTGATTTAGACTTTGCACTGCGAAGTCCTAACCCTGGTGATATATTTGAAA 7125 7126 AGAAAATCTTACCGAAATTTCCTCCAGTTTTTTATGAGTGGTTTGTTGAATCTTTCCCAGAACCAAATAATTGGG 7200 7201 TTACTAGTAGACAAAACTATTGCCGAACTTTAGCAGTAATGTCAATAGTTGGCTACGTTTTGGGTTTGGGAGATC 7275 7276 GCCATGGCGAAAACATATTGTTTGATGAATTTACAGGTGAAGCTATCCATGTCGATTTCAACTGTCTTTTTGATA 7350 7351 AAGGTCTTACTTTTGAAAAACCTGAAAAGGTGCCGTTCAGATTAACTCATAATATGGTAGATGCAATGGGTCCGA 7425 7426 CAGGTTATGAAGGGGGTTTCAGGAAAGCTAGCGAAATAACGATGCGGCTTCTTCGCTCAAACCAAGATACATTGA 7500 7501 TGAGCGTACTAGAGTCTTTCCTACATGATCCTTTAGTCGAGTGGAATAGAAAGAAGTCGTCAAGCAAGTACCCGA 7575 7576 ATAATGAAGCAAATGAAGTTTTGGATATAATTCGCAAAAAATTTCAAGGCTTTATGCCAGGGGAGACGATACCTT 7650 7651 TATCTATTGAAGGGCAAATTCAAGAATTGATCAAATCTGCTGTCAACCCAAAAAACCTGGTAGAAATGTACATTG 7725 7726 GTTGGGCTGCTTATTTCTAGCATTTTACTAACAAAAATTTCAATGAACAAGCTACCCATTATTAAACTTATGATT 7800 7801 TGAATCGAAGATATTTTATTTATTAATCCGATGAAGAATTCTCGCTGAGTTGTTCAATTTCTTGTAATTTTCCTT 7875 7876 CCATTTCTAAATCGTCGATTCGCTTAAATAGGGCACTGGCTTTTTGTGCATTTTTCTCTCGTAAAGCAGCTTCTG 7950 7951 ATTGAAAAAAAGCTATATCTGTTTCTGAGTCATCATCCGAATCAACAATATATTTTGCAGATCGACCTGCAG    8022 In italics, sequenced by Seaton, et al. On Bold are those bases deleted in Seaton et al. (2499, 22501, 2507, 2509) Underlined are the two bases either side of a single C insert (5918/5919) in Seaton et al. (i.e., incorrect base not shown, but the one residue either side is) Sequence ID No. 4: rad3 protein    1 MSQHAKRKAGSLDLSPRGLDDRQAFGQLLKEVLALDKEHELGRSNSLPSMTSELVEVLIEVGLLAFKHDDSKSEF   75   76 ISPKMLLEAHLSLQALMLILKRSPTVLREIKSSVTLLDWILPRTISLFADIRFIKLFDSLKEFHKLIYQLISEKS  150  151 FLWDLYASFMRYWKYYITNVSSIVLQITNATFPYKMPSPNSQPLQSISPNYPTHREDKFDLLIINIEEACTFFFE  225  226 SAHFFAQCSYLKKSNFPSPPLFTAWTWIKPCFFNFVILLKRISIGDSQLFLHLHSRIVQTLCCFSLNFIYHGLPI  300  301 CEKSKHILMSSINLTLGSLKKTYTVANTAISLFFLSLFVLPKTVAGLFYPFGVSLLSDFKVLEQLEPDSDLKKAI  375  376 ILFKCRYQSSEIDQTTLRAFGEICTGKLENTLFSNSELMLFLLHYLSLDNDLSNILKVDFQNGHNICTFAKWCIN  450  451 NNLDEPSNLKHFREMLDYYSSHNVTISEDDLKNFSLVLCTHVAKVNEKTNSIFRTYEVHGCEVCNSFCKKFDERS  525  526 PFKIPYHELFCALLKNPDIISSSVKQSLLLDGFFRWSQHCSNFNKESMLSLREFIMKALASTSRCLRVVAAKVLP  600  601 IFIKGPNNLDIVEFHKESKALIFNTLKILAVENTAILETVILSWISLSRVVEEEELHFVLLEVISSVINSGIFYQ  675  676 GIGLSALQQIASTRHISVWQKKSPYWPTVSVAIVQGMGKKPNIASLFAQLMNISEGDFLIRTQAYTLPFLVLTKN  750  751 KALIVRIAELSQSDVATLCLTNMHKILASLLIIDHPNLEESVMLLLSLATSDFEKVDLTSLLRSDPISITVELLQ  825  826 LYQNDVPHEKIENALRKVAMIVSQVVNDEDLSNKELLYDFFNNHILGILAEFSNILNDLKGKTSINEKIKTIVGI  900  901 EKMLSLCGGAVKLGLPQILSNLQSAFQNEHLRFYAIKAWFSLILATKEPEYSSIAGLSLVILPPLFPYLEPQEAE  975  976 LVIQIFDFISSDTHKCLQGLKWAIPTSLDSACFSLKAKEIFCSLQNEDFYSELQSIIKCLTNENEPVCYLGLQKL 1050 1051 ELFFQAKVDELHDTLNLDISNEVLDQLLRCLLDCCVKYASTNMQISYLAAKNLGELGAIDPSRAKAQHIIKETVV 1125 1126 LDNFENGEESLKFILDFMQSQLIPAFLVTTDTKAQGFLAYALQEFLKLGGFKSAVINKKKGLTVVTEHWMSLPDL 1200 1201 SKRVLIPFLTSKYHLTPIPKIDIRYPIYKENVTIHTWMQLFSLKLMEYAHSQNAEKIFGICSKVVKDQEVNIPCF 1275 1276 LLPFLVLNVILTESELEVNKVIEEFQLVINQPGPDGLNSVGQQRYTSFVDVFFKIVDYLNKWLRMRKKRNWDRRS 1350 1351 AIARKENRYMSVEDATSRESSISKVESFLSRFPSKTLGIVSLNCGFHARALFYWEQHIRNATAPYAALESDYRVL 1425 1426 QEIYAGIDDPDEIEAVSLNNFHDYSFDQQLLHENSGTWDSALSCYEIIIQKDPENKKAKIGLLNSMLQSGHYESL 1500 1501 VLSLDSFIINDNHEYSKMLNLGIEASWRSLSIDSLKKCLSKSNLESFEAKLGSIFYQYLRKDSFAELTERLQPLY 1575 1576 VDAATAIANTGAHSAYDCYDILSKLHAINDFSRIAETDGIVSDNLDIVLRRRLSQVAPYGKFKHQILSTHLVGYE 1650 1651 KFENTKKTAEIYLEIARISRKNGQFQRAFNAILKAMDLDKPLATIEHAQWWWHQGQHRKAISELNFSLNNNMFDL 1725 1726 VDEHEERPKNRKETLGNPLKGKVFLKLTKWLGKAGQLGLKDLETYYHKAVEIYSECENTHYYLGHHRVLMYEEEQ 1800 1801 KLPVNEQSERFLSGELVTRIINEFGRSLYYGTNHIYESMPKLLTLWLDFGAEELRLSKDDGEKYFREHIISSRKK 1875 1876 SLELMNSNVCRLSMKIPQYFFLVALSQMISRVCHPNNKVYKILEHIIANVVASYPGETLWQLMATIKSTSQKRSL 1950 1951 RGKSILNVLHSRKLSMSSKVDIKALSQSAILITEKLINLCNTRINSKSVKMSLKDHFRLSFDDPVDLVIPAKSFL 2025 2026 DITLPAKDANRASHYPFPKTQPTLLKFEDEVDIMNSLQKPRKVYVRGTDGNLYPFLCKPKDDLRKDARLMEFNNL 2100 2101 ICKILRKDQEANRRNLCIRTYVVIPLNEECGFIEWVNHTRPFREILLKSYRQKNIPISYQEIKVDLDFALRSPNP 2175 2176 GDIFEKKILPKFPPVFYEWFVESFPEPNNWVTSRQNYCRTLAVMSIVGYVLGLGDRHGENILFDEFTGEAIHVDF 2250 2251 NCLFDKGLTFEKPEKVPFRLTHNMVDAMGPTGYEGGFRKASEITMRLLRSNQDTLMSVLESFLHDPLVEWNRKKS 2325 2326 SSKYPNNEANEVLDIIRKKFQGFMPGETIPLSIEGQIQELIKSAVNPKNLVEMYIGWAAYF               2386

                   #             SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 14 <210> SEQ ID NO 1 <211> LENGTH: 8239 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (80)..(8011) <400> SEQUENCE: 1 gcgctcttcc ggcagcggta cgtttggaga cgccgggaac ccgcgttggc gt #ggttgact     60 agtgcctcgc agcctcagc atg ggg gaa cat ggc ctg gag # ctg gct tcc atg     112                    #   Met Gly Glu His Gly Leu Glu Leu Ala # Ser Met                    #     1              #  5                 #  10 atc ccc gcc ctg cgg gag ctg ggc agt gcc ac #a cca gag gaa tat aat      160 Ile Pro Ala Leu Arg Glu Leu Gly Ser Ala Th #r Pro Glu Glu Tyr Asn              15      #             20      #             25 aca gtt gta cag aag cca aga caa att ctg tg #t caa ttc att gac cgg      208 Thr Val Val Gln Lys Pro Arg Gln Ile Leu Cy #s Gln Phe Ile Asp Arg          30          #         35          #         40 ata ctt aca gat gta aat gtt gtt gct gta ga #a ctt gta aag aaa act      256 Ile Leu Thr Asp Val Asn Val Val Ala Val Gl #u Leu Val Lys Lys Thr      45              #     50              #     55 gac tct cag cca acc tcc gtg atg ttg ctt ga #t ttc atc cag cat atc      304 Asp Ser Gln Pro Thr Ser Val Met Leu Leu As #p Phe Ile Gln His Ile  60                  # 65                  # 70                  # 75 atg aaa tcc tcc cca ctt atg ttt gta aat gt #g agt gga agc cat gag      352 Met Lys Ser Ser Pro Leu Met Phe Val Asn Va #l Ser Gly Ser His Glu                  80  #                 85  #                 90 cgc aaa ggc agt tgt att gaa ttc agt aat tg #g atc ata acg aga ctt      400 Arg Lys Gly Ser Cys Ile Glu Phe Ser Asn Tr #p Ile Ile Thr Arg Leu              95      #            100      #            105 ctg cgg att gca gca act ccc tcc tgt cat tt #g tta cac aag aaa atc      448 Leu Arg Ile Ala Ala Thr Pro Ser Cys His Le #u Leu His Lys Lys Ile         110           #       115           #       120 tgt gaa gtc atc tgt tca tta tta ttt ctt tt #t aaa agc aag agt cct      496 Cys Glu Val Ile Cys Ser Leu Leu Phe Leu Ph #e Lys Ser Lys Ser Pro     125               #   130               #   135 gct att ttt ggg gta ctc aca aaa gaa tta tt #a caa ctt ttt gaa gac      544 Ala Ile Phe Gly Val Leu Thr Lys Glu Leu Le #u Gln Leu Phe Glu Asp 140                 1 #45                 1 #50                 1 #55 ttg gtt tac ctc cat aga aga aat gtg atg gg #t cat gct gtg gaa tgg      592 Leu Val Tyr Leu His Arg Arg Asn Val Met Gl #y His Ala Val Glu Trp                 160   #               165   #               170 cca gtg gtc atg agc cga ttt tta agt caa tt #a gat gaa cac atg gga      640 Pro Val Val Met Ser Arg Phe Leu Ser Gln Le #u Asp Glu His Met Gly             175       #           180       #           185 tat tta caa tca gct cct ttg cag ttg atg ag #t atg caa aat tta gaa      688 Tyr Leu Gln Ser Ala Pro Leu Gln Leu Met Se #r Met Gln Asn Leu Glu         190           #       195           #       200 ttt att gaa gtc act tta tta atg gtt ctt ac #t cgt att att gca att      736 Phe Ile Glu Val Thr Leu Leu Met Val Leu Th #r Arg Ile Ile Ala Ile     205               #   210               #   215 gtg ttt ttt aga agg caa gaa ctc tta ctt tg #g cag ata ggt tgt gtt      784 Val Phe Phe Arg Arg Gln Glu Leu Leu Leu Tr #p Gln Ile Gly Cys Val 220                 2 #25                 2 #30                 2 #35 ctg cta gag tat ggt agt cca aaa att aaa tc #c cta gca att agc ttt      832 Leu Leu Glu Tyr Gly Ser Pro Lys Ile Lys Se #r Leu Ala Ile Ser Phe                 240   #               245   #               250 tta aca gaa ctt ttt cag ctt gga gga cta cc #a gca caa cca gct agc      880 Leu Thr Glu Leu Phe Gln Leu Gly Gly Leu Pr #o Ala Gln Pro Ala Ser             255       #           260       #           265 act ttt ttc agc tca ttt ttg gaa tta tta aa #a cac ctt gta gaa atg      928 Thr Phe Phe Ser Ser Phe Leu Glu Leu Leu Ly #s His Leu Val Glu Met         270           #       275           #       280 gat act gac caa ttg aaa ctc tat gaa gag cc #a tta tca aag ctg ata      976 Asp Thr Asp Gln Leu Lys Leu Tyr Glu Glu Pr #o Leu Ser Lys Leu Ile     285               #   290               #   295 aag aca cta ttt ccc ttt gaa gca gaa gct ta #t aga aat att gaa cct     1024 Lys Thr Leu Phe Pro Phe Glu Ala Glu Ala Ty #r Arg Asn Ile Glu Pro 300                 3 #05                 3 #10                 3 #15 gtc tat tta aat atg ctg ctg gaa aaa ctc tg #t gtc atg ttt gaa gac     1072 Val Tyr Leu Asn Met Leu Leu Glu Lys Leu Cy #s Val Met Phe Glu Asp                 320   #               325   #               330 ggt gtg ctc atg cgg ctt aag tct gat ttg ct #a aaa gca gct ttg tgc     1120 Gly Val Leu Met Arg Leu Lys Ser Asp Leu Le #u Lys Ala Ala Leu Cys             335       #           340       #           345 cat tta ctg cag tat ttc ctt aaa ttt gtg cc #a gct ggg tat gaa tct     1168 His Leu Leu Gln Tyr Phe Leu Lys Phe Val Pr #o Ala Gly Tyr Glu Ser         350           #       355           #       360 gct tta caa gtc agg aag gtc tat gtg aga aa #t att tgt aaa gct ctt     1216 Ala Leu Gln Val Arg Lys Val Tyr Val Arg As #n Ile Cys Lys Ala Leu     365               #   370               #   375 ttg gat gtg ctt gga att gag gta gat gca ga #g tac ttg ttg ggc cca     1264 Leu Asp Val Leu Gly Ile Glu Val Asp Ala Gl #u Tyr Leu Leu Gly Pro 380                 3 #85                 3 #90                 3 #95 ctt tat gca gct ttg aaa atg gaa agt atg ga #a atc att gag gag att     1312 Leu Tyr Ala Ala Leu Lys Met Glu Ser Met Gl #u Ile Ile Glu Glu Ile                 400   #               405   #               410 caa tgc caa act caa cag gaa aac ctc agc ag #t aat agt gat gga ata     1360 Gln Cys Gln Thr Gln Gln Glu Asn Leu Ser Se #r Asn Ser Asp Gly Ile             415       #           420       #           425 tca ccc aaa agg cgt cgt ctc agc tcg tct ct #a aac cct tct aaa aga     1408 Ser Pro Lys Arg Arg Arg Leu Ser Ser Ser Le #u Asn Pro Ser Lys Arg         430           #       435           #       440 gca cca aaa cag act gag gaa att aaa cat gt #g gac atg aac caa aag     1456 Ala Pro Lys Gln Thr Glu Glu Ile Lys His Va #l Asp Met Asn Gln Lys     445               #   450               #   455 agc ata tta tgg agt gca ctg aaa cag aaa gc #t gaa tcc ctt cag att     1504 Ser Ile Leu Trp Ser Ala Leu Lys Gln Lys Al #a Glu Ser Leu Gln Ile 460                 4 #65                 4 #70                 4 #75 tcc ctt gaa tac agt ggc cta aag aat cct gt #t att gag atg tta gaa     1552 Ser Leu Glu Tyr Ser Gly Leu Lys Asn Pro Va #l Ile Glu Met Leu Glu                 480   #               485   #               490 gga att gct gtt gtc tta caa ctg act gct ct #g tgt act gtt cat tgt     1600 Gly Ile Ala Val Val Leu Gln Leu Thr Ala Le #u Cys Thr Val His Cys             495       #           500       #           505 tct cat caa aac atg aac tgc cgt act ttc aa #g gac tgt caa cat aaa     1648 Ser His Gln Asn Met Asn Cys Arg Thr Phe Ly #s Asp Cys Gln His Lys         510           #       515           #       520 tcc aag aag aaa cct tct gta gtg ata act tg #g atg tca ttg gat ttt     1696 Ser Lys Lys Lys Pro Ser Val Val Ile Thr Tr #p Met Ser Leu Asp Phe     525               #   530               #   535 tac aca aaa gtg ctt aag agc tgt aga agt tt #g tta gaa tct gtt cag     1744 Tyr Thr Lys Val Leu Lys Ser Cys Arg Ser Le #u Leu Glu Ser Val Gln 540                 5 #45                 5 #50                 5 #55 aaa ctg gac ctg gag gca acc att gat aag gt #g gtg aaa att tat gat     1792 Lys Leu Asp Leu Glu Ala Thr Ile Asp Lys Va #l Val Lys Ile Tyr Asp                 560   #               565   #               570 gct ttg att tat atg caa gta aac agt tca tt #t gaa gat cat atc ctg     1840 Ala Leu Ile Tyr Met Gln Val Asn Ser Ser Ph #e Glu Asp His Ile Leu             575       #           580       #           585 gaa gat tta tgt ggt atg ctc tca ctt cca tg #g att tat tcc cat tct     1888 Glu Asp Leu Cys Gly Met Leu Ser Leu Pro Tr #p Ile Tyr Ser His Ser         590           #       595           #       600 gat gat ggc tgt tta aag ttg acc aca ttt gc #c gct aat ctt cta aca     1936 Asp Asp Gly Cys Leu Lys Leu Thr Thr Phe Al #a Ala Asn Leu Leu Thr     605               #   610               #   615 tta agc tgt agg att tca gat agc tat tca cc #a cag gca caa tca cga     1984 Leu Ser Cys Arg Ile Ser Asp Ser Tyr Ser Pr #o Gln Ala Gln Ser Arg 620                 6 #25                 6 #30                 6 #35 tgt gtg ttt ctt ctg act ctg ttt cca aga ag #a ata ttc ctt gag tgg     2032 Cys Val Phe Leu Leu Thr Leu Phe Pro Arg Ar #g Ile Phe Leu Glu Trp                 640   #               645   #               650 aga aca gca gtt tac aac tgg gcc ctg cag ag #c tcc cat gaa gta atc     2080 Arg Thr Ala Val Tyr Asn Trp Ala Leu Gln Se #r Ser His Glu Val Ile             655       #           660       #           665 cgg gct agt tgt gtt agt gga ttt ttt atc tt #a ttg cag cag cag aat     2128 Arg Ala Ser Cys Val Ser Gly Phe Phe Ile Le #u Leu Gln Gln Gln Asn         670           #       675           #       680 tct tgt aac aga gtt ccc aag att ctt ata ga #t aaa gtc aaa gat gat     2176 Ser Cys Asn Arg Val Pro Lys Ile Leu Ile As #p Lys Val Lys Asp Asp     685               #   690               #   695 tct gac att gtc aag aaa gaa ttt gct tct at #a ctt ggt caa ctt gtc     2224 Ser Asp Ile Val Lys Lys Glu Phe Ala Ser Il #e Leu Gly Gln Leu Val 700                 7 #05                 7 #10                 7 #15 tgt act ctt cac ggc atg ttt tat ctg aca ag #t tct tta aca gaa cct     2272 Cys Thr Leu His Gly Met Phe Tyr Leu Thr Se #r Ser Leu Thr Glu Pro                 720   #               725   #               730 ttc tct gaa cac gga cat gtg gac ctc ttc tg #t agg aac ttg aaa gcc     2320 Phe Ser Glu His Gly His Val Asp Leu Phe Cy #s Arg Asn Leu Lys Ala             735       #           740       #           745 act tct caa cat gaa tgt tca tct tct caa ct #a aaa gct tct gtc tgc     2368 Thr Ser Gln His Glu Cys Ser Ser Ser Gln Le #u Lys Ala Ser Val Cys         750           #       755           #       760 aag cca ttc ctt ttc cta ctg aaa aaa aaa at #a cct agt cca gta aaa     2416 Lys Pro Phe Leu Phe Leu Leu Lys Lys Lys Il #e Pro Ser Pro Val Lys     765               #   770               #   775 ctt gct ttc ata gat aat cta cat cat ctt tg #t aag cat ctt gat ttt     2464 Leu Ala Phe Ile Asp Asn Leu His His Leu Cy #s Lys His Leu Asp Phe 780                 7 #85                 7 #90                 7 #95 aga gaa gat gaa aca gat gta aaa gca gtt ct #t gga act tta tta aat     2512 Arg Glu Asp Glu Thr Asp Val Lys Ala Val Le #u Gly Thr Leu Leu Asn                 800   #               805   #               810 tta atg gaa gat cca gac aaa gat gtt aga gt #g gct ttt agt gga aat     2560 Leu Met Glu Asp Pro Asp Lys Asp Val Arg Va #l Ala Phe Ser Gly Asn             815       #           820       #           825 atc aag cac ata ttg gaa tcc ttg gac tct ga #a gat gga ttt ata aag     2608 Ile Lys His Ile Leu Glu Ser Leu Asp Ser Gl #u Asp Gly Phe Ile Lys         830           #       835           #       840 gag ctt ttt gtc tta aga atg aag gaa gca ta #t aca cat gcc caa ata     2656 Glu Leu Phe Val Leu Arg Met Lys Glu Ala Ty #r Thr His Ala Gln Ile     845               #   850               #   855 tca aga aat aat gag ctg aag gat acc ttg at #t ctt aca aca ggg gat     2704 Ser Arg Asn Asn Glu Leu Lys Asp Thr Leu Il #e Leu Thr Thr Gly Asp 860                 8 #65                 8 #70                 8 #75 att gga agg gcc gca aaa gga gat ttg gta cc #a ttt gca ctc tta cac     2752 Ile Gly Arg Ala Ala Lys Gly Asp Leu Val Pr #o Phe Ala Leu Leu His                 880   #               885   #               890 tta ttg cat tgt ttg tta tcc aag tca gca tc #t gtc tct gga gca gca     2800 Leu Leu His Cys Leu Leu Ser Lys Ser Ala Se #r Val Ser Gly Ala Ala             895       #           900       #           905 tac aca gaa att aga gct ctg gtt gca gct aa #a agt gtt aaa ctg caa     2848 Tyr Thr Glu Ile Arg Ala Leu Val Ala Ala Ly #s Ser Val Lys Leu Gln         910           #       915           #       920 agt ttt ttc agc cag tat aag aaa ccc atc tg #t cag ttt ttg gta gaa     2896 Ser Phe Phe Ser Gln Tyr Lys Lys Pro Ile Cy #s Gln Phe Leu Val Glu     925               #   930               #   935 tcc ctt cac tct agt cag atg aca gca ctt cc #g aat act cca tgc cag     2944 Ser Leu His Ser Ser Gln Met Thr Ala Leu Pr #o Asn Thr Pro Cys Gln 940                 9 #45                 9 #50                 9 #55 aat gct gac gtg cga aaa caa gat gtg gct ca #c cag aga gaa atg gct     2992 Asn Ala Asp Val Arg Lys Gln Asp Val Ala Hi #s Gln Arg Glu Met Ala                 960   #               965   #               970 tta aat acg ttg tct gaa att gcc aac gtt tt #c gac ttt cct gat ctt     3040 Leu Asn Thr Leu Ser Glu Ile Ala Asn Val Ph #e Asp Phe Pro Asp Leu             975       #           980       #           985 aat cgt ttt ctt act agg aca tta caa gtt ct #a cta cct  gat ctt gct    3088 Asn Arg Phe Leu Thr Arg Thr Leu Gln Val Le #u Leu Pro  Asp Leu Ala         990           #       995           #       1000 gcc aaa  gca agc cct gca gct  tct gct c #tc att cga  act tta gga aaa  3136 Ala Lys  Ala Ser Pro Ala Ala  Ser Ala L #eu Ile Arg  Thr Leu Gly Lys     1005              #    1010              #    1015 caa  tta aat gtc aat cgt  aga gag att t #ta ata  aac aac ttc aaa tat  3184 Gln  Leu Asn Val Asn Arg  Arg Glu Ile L #eu Ile  Asn Asn Phe Lys Tyr 1020                1025 #                 103 #0                 10 #35 att ttt tct cat ttg  gtc tgt tct tgt tcc  # aaa gat gaa tta gaa  cgt  3232 Ile Phe Ser His Leu  Val Cys Ser Cys Ser  # Lys Asp Glu Leu Glu  Arg                 1040  #                1045  #                1050 gcc ctt cat tat  ctg aag aat gaa aca  g #aa att gaa ctg ggg  agc ctg  3280 Ala Leu His Tyr  Leu Lys Asn Glu Thr  G #lu Ile Glu Leu Gly  Ser Leu             1055      #            1060      #            1065 ttg aga caa  gat ttc caa gga ttg  cat a #at gaa tta ttg  ctg cgt att  3328 Leu Arg Gln  Asp Phe Gln Gly Leu  His A #sn Glu Leu Leu  Leu Arg Ile         1070          #        1075          #        1080 gga gaa  cac tat caa cag gtt  ttt aat g #gt ttg tca  ata ctt gcc tca  3376 Gly Glu  His Tyr Gln Gln Val  Phe Asn G #ly Leu Ser  Ile Leu Ala Ser     1085              #    1090              #    1095 ttt  gca tcc agt gat gat  cca tat cag g #gc ccg  aga gat atc ata tca  3424 Phe  Ala Ser Ser Asp Asp  Pro Tyr Gln G #ly Pro  Arg Asp Ile Ile Ser 1100                1105 #                 111 #0                 11 #15 cct gaa ctg atg gct  gat tat tta caa ccc  # aaa ttg ttg ggc att  ttg  3472 Pro Glu Leu Met Ala  Asp Tyr Leu Gln Pro  # Lys Leu Leu Gly Ile  Leu                 1120  #                1125  #                1130 gct ttt ttt aac  atg cag tta ctg agc  t #ct agt gtt ggc att  gaa gat  3520 Ala Phe Phe Asn  Met Gln Leu Leu Ser  S #er Ser Val Gly Ile  Glu Asp             1135      #            1140      #            1145 aag aaa atg  gcc ttg aac agt ttg  atg t #ct ttg atg aag  tta atg gga  3568 Lys Lys Met  Ala Leu Asn Ser Leu  Met S #er Leu Met Lys  Leu Met Gly         1150          #        1155          #        1160 ccc aaa  cat gtc agt tct gtg  agg gtg a #ag atg atg  acc aca ctg aga  3616 Pro Lys  His Val Ser Ser Val  Arg Val L #ys Met Met  Thr Thr Leu Arg     1165              #    1170              #    1175 act  ggc ctt cga ttc aag  gat gat ttt c #ct gaa  ttg tgt tgc aga gct  3664 Thr  Gly Leu Arg Phe Lys  Asp Asp Phe P #ro Glu  Leu Cys Cys Arg Ala 1180                  #1185                  #1190                  #1195 tgg gac tgc ttt gtt  cgc tgc ctg gat cat  # gct tgt ctg ggc tcc  ctt  3712 Trp Asp Cys Phe Val  Arg Cys Leu Asp His  # Ala Cys Leu Gly Ser  Leu                 1200  #                1205  #                1210 ctc agt cat gta  ata gta gct ttg tta  c #ct ctt ata cac atc  cag cct  3760 Leu Ser His Val  Ile Val Ala Leu Leu  P #ro Leu Ile His Ile  Gln Pro             1215      #            1220      #            1225 aaa gaa act  gca gct atc ttc cac  tac c #tc ata att gaa  aac agg gat  3808 Lys Glu Thr  Ala Ala Ile Phe His  Tyr L #eu Ile Ile Glu  Asn Arg Asp         1230          #        1235          #        1240 gct gtg  caa gat ttt ctt cat  gaa ata t #at ttt tta  cct gat cat cca  3856 Ala Val  Gln Asp Phe Leu His  Glu Ile T #yr Phe Leu  Pro Asp His Pro     1245              #    1250              #    1255 gaa  tta aaa aag ata aaa  gcc gtt ctc c #ag gaa  tac aga aag gag acc  3904 Glu  Leu Lys Lys Ile Lys  Ala Val Leu G #ln Glu  Tyr Arg Lys Glu Thr 1260                  #1265                  #1270                  #1275 tct gag agc act gat  ctt cag aca act ctt  # cag ctc tct atg aag  gcc  3952 Ser Glu Ser Thr Asp  Leu Gln Thr Thr Leu  # Gln Leu Ser Met Lys  Ala                 1280  #                1285  #                1290 att caa cat gaa  aat gtc gat gtt cgt  a #tt cat gct ctt aca  agc ttg  4000 Ile Gln His Glu  Asn Val Asp Val Arg  I #le His Ala Leu Thr  Ser Leu             1295      #            1300      #            1305 aag gaa acc  ttg tat aaa aat cag  gaa a #aa ctg ata aag  tat gca aca  4048 Lys Glu Thr  Leu Tyr Lys Asn Gln  Glu L #ys Leu Ile Lys  Tyr Ala Thr         1310          #        1315          #        1320 gac agt  gaa aca gta gaa cct  att atc t #ca cag ttg  gtg aca gtg ctt  4096 Asp Ser  Glu Thr Val Glu Pro  Ile Ile S #er Gln Leu  Val Thr Val Leu     1325              #    1330              #    1335 ttg  aaa ggt tgc caa gat  gca aac tct c #aa gct  cgg ttg ctc tgt ggg  4144 Leu  Lys Gly Cys Gln Asp  Ala Asn Ser G #ln Ala  Arg Leu Leu Cys Gly 1340                  #1345                  #1350                  #1355 gaa tgt tta ggg gaa  ttg ggg gcg ata gat  # cca ggt cga tta gat  ttc  4192 Glu Cys Leu Gly Glu  Leu Gly Ala Ile Asp  # Pro Gly Arg Leu Asp  Phe                 1360  #                1365  #                1370 tca aca act gaa  act caa gga aaa gat  t #tt aca ttt gtg act  gga gta  4240 Ser Thr Thr Glu  Thr Gln Gly Lys Asp  P #he Thr Phe Val Thr  Gly Val             1375      #            1380      #            1385 gaa gat tca  agc ttt gcc tat gga  tta t #tg atg gag cta  aca aga gct  4288 Glu Asp Ser  Ser Phe Ala Tyr Gly  Leu L #eu Met Glu Leu  Thr Arg Ala         1390          #        1395          #        1400 tac ctt  gcg tat gct gat aat  agc cga g #ct caa gat  tca gct gcc tat  4336 Tyr Leu  Ala Tyr Ala Asp Asn  Ser Arg A #la Gln Asp  Ser Ala Ala Tyr     1405              #    1410              #    1415 gcc  att cag gag ttg ctt  tct att tat g #ac tgt  aga gag atg gag acc  4384 Ala  Ile Gln Glu Leu Leu  Ser Ile Tyr A #sp Cys  Arg Glu Met Glu Thr 1420                  #1425                  #1430                  #1435 aac ggc cca ggt cac  caa ttg tgg agg aga  # ttt cct gag cat gtt  cgg  4432 Asn Gly Pro Gly His  Gln Leu Trp Arg Arg  # Phe Pro Glu His Val  Arg                 1440  #                1445  #                1450 gaa ata cta gaa  cct cat cta aat acc  a #ga tac aag agt tct  cag aag  4480 Glu Ile Leu Glu  Pro His Leu Asn Thr  A #rg Tyr Lys Ser Ser  Gln Lys             1455      #            1460      #            1465 tca acc gat  tgg tct gga gta aag  aag c #ca att tac tta  agt aaa ttg  4528 Ser Thr Asp  Trp Ser Gly Val Lys  Lys P #ro Ile Tyr Leu  Ser Lys Leu         1470          #        1475          #        1480 ggt agt  aac ttt gca gaa tgg  tca gca t #ct tgg gca  ggt tat ctt att  4576 Gly Ser  Asn Phe Ala Glu Trp  Ser Ala S #er Trp Ala  Gly Tyr Leu Ile     1485              #    1490              #    1495 aca  aag gtt cga cat gat  ctt gcc agt a #aa att  ttc acc tgc tgt agc  4624 Thr  Lys Val Arg His Asp  Leu Ala Ser L #ys Ile  Phe Thr Cys Cys Ser 1500                  #1505                  #1510                  #1515 att atg atg aag cat  gat ttc aaa gtg acc  # atc tat ctt ctt cca  cat  4672 Ile Met Met Lys His  Asp Phe Lys Val Thr  # Ile Tyr Leu Leu Pro  His                 1520  #                1525  #                1530 att ctg gtg tat  gtc tta ctg ggt tgt  a #at caa gaa gat cag  cag gag  4720 Ile Leu Val Tyr  Val Leu Leu Gly Cys  A #sn Gln Glu Asp Gln  Gln Glu             1535      #            1540      #            1545 gtt tat gca  gaa att atg gca gtt  cta a #ag cat gac gat  cag cat acc  4768 Val Tyr Ala  Glu Ile Met Ala Val  Leu L #ys His Asp Asp  Gln His Thr         1550          #        1555          #        1560 ata aat  acc caa gac att gca  tct gat c #tg tgt caa  ctc agt aca cag  4816 Ile Asn  Thr Gln Asp Ile Ala  Ser Asp L #eu Cys Gln  Leu Ser Thr Gln     1565              #    1570              #    1575 act  gtg ttc tcc atg ctt  gac cat ctc a #ca cag  tgg gca agg cac aaa  4864 Thr  Val Phe Ser Met Leu  Asp His Leu T #hr Gln  Trp Ala Arg His Lys 1580                  #1585                  #1590                  #1595 ttt cag gca ctg aaa  gct gag aaa tgt cca  # cac agc aaa tca aac  aga  4912 Phe Gln Ala Leu Lys  Ala Glu Lys Cys Pro  # His Ser Lys Ser Asn  Arg                 1600  #                1605  #                1610 aat aag gta gac  tca atg gta tct act  g #tg gat tat gaa gac  tat cag  4960 Asn Lys Val Asp  Ser Met Val Ser Thr  V #al Asp Tyr Glu Asp  Tyr Gln             1615      #            1620      #            1625 agt gta acc  cgt ttt cta gac ctc  ata c #cc cag gat act  ctg gca gta  5008 Ser Val Thr  Arg Phe Leu Asp Leu  Ile P #ro Gln Asp Thr  Leu Ala Val         1630          #        1635          #        1640 gct tcc  ttt cgc tcc aaa gca  tac aca c #ga gct gta  atg cac ttt gaa  5056 Ala Ser  Phe Arg Ser Lys Ala  Tyr Thr A #rg Ala Val  Met His Phe Glu     1645              #    1650              #    1655 tca  ttt att aca gaa aag  aag caa aat a #tt cag  gaa cat ctt gga ttt  5104 Ser  Phe Ile Thr Glu Lys  Lys Gln Asn I #le Gln  Glu His Leu Gly Phe 1660                  #1665                  #1670                  #1675 tta cag aaa ttg tat  gct gct atg cat gaa  # cct gat gga gtg gcc  gga  5152 Leu Gln Lys Leu Tyr  Ala Ala Met His Glu  # Pro Asp Gly Val Ala  Gly                 1680  #                1685  #                1690 gtc agt gca att  aga aag gca gaa cca  t #ct cta aaa gaa cag  atc ctt  5200 Val Ser Ala Ile  Arg Lys Ala Glu Pro  S #er Leu Lys Glu Gln  Ile Leu             1695      #            1700      #            1705 gaa cat gaa  agc ctt ggc ttg ctg  agg g #at gcc act gct  tgt tat gac  5248 Glu His Glu  Ser Leu Gly Leu Leu  Arg A #sp Ala Thr Ala  Cys Tyr Asp         1710          #        1715          #        1720 agg gct  att cag cta gaa cca  gac cag a #tc att cat  tat cat ggt gta  5296 Arg Ala  Ile Gln Leu Glu Pro  Asp Gln I #le Ile His  Tyr His Gly Val     1725              #    1730              #    1735 gta  aag tcc atg tta ggt  ctt ggt cag c #tg tct  act gtt atc act cag  5344 Val  Lys Ser Met Leu Gly  Leu Gly Gln L #eu Ser  Thr Val Ile Thr Gln 1740                  #1745                  #1750                  #1755 gtg aat gga gtg cat  gct aac agg tcc gag  # tgg aca gat gaa tta  aac  5392 Val Asn Gly Val His  Ala Asn Arg Ser Glu  # Trp Thr Asp Glu Leu  Asn                 1760  #                1765  #                1770 acg tac aga gtg  gaa gca gct tgg aaa  t #tg tca cag tgg gat  ttg gtg  5440 Thr Tyr Arg Val  Glu Ala Ala Trp Lys  L #eu Ser Gln Trp Asp  Leu Val             1775      #            1780      #            1785 gaa aac tat  ttg gca gca gat gga  aaa t #ct aca aca tgg  agt gtc aga  5488 Glu Asn Tyr  Leu Ala Ala Asp Gly  Lys S #er Thr Thr Trp  Ser Val Arg         1790          #        1795          #        1800 ctg gga  cag cta tta tta tca  gcc aaa a #aa aga gat  atc aca gct ttt  5536 Leu Gly  Gln Leu Leu Leu Ser  Ala Lys L #ys Arg Asp  Ile Thr Ala Phe     1805              #    1810              #    1815 tat  gac tca ctg aaa cta  gtg aga gca g #aa caa  att gta cct ctt tca  5584 Tyr  Asp Ser Leu Lys Leu  Val Arg Ala G #lu Gln  Ile Val Pro Leu Ser 1820                  #1825                  #1830                  #1835 gct gca agc ttt gaa  aga ggc tcc tac caa  # cga gga tat gaa tat  att  5632 Ala Ala Ser Phe Glu  Arg Gly Ser Tyr Gln  # Arg Gly Tyr Glu Tyr  Ile                 1840  #                1845  #                1850 gtg aga ttg cac  atg tta tgt gag ttg  g #ag cat agc atc aaa  cca ctt  5680 Val Arg Leu His  Met Leu Cys Glu Leu  G #lu His Ser Ile Lys  Pro Leu             1855      #            1860      #            1865 ttc cag cat  tct cca ggt gac agt  tct c #aa gaa gat tct  cta aac tgg  5728 Phe Gln His  Ser Pro Gly Asp Ser  Ser G #ln Glu Asp Ser  Leu Asn Trp         1870          #        1875          #        1880 gta gct  cga cta gaa atg acc  cag aat t #cc tac aga  gcc aag gac cct  5776 Val Ala  Arg Leu Glu Met Thr  Gln Asn S #er Tyr Arg  Ala Lys Asp Pro     1885              #    1890              #    1895 atc  ctg gct ctc cgg agg  gct tta cta a #gc ctc  aac aaa aga cca gat  5824 Ile  Leu Ala Leu Arg Arg  Ala Leu Leu S #er Leu  Asn Lys Arg Pro Asp 1900                  #1905                  #1910                  #1915 tac aat gaa atg gtt  gga gaa tgc tgg ctg  # cag agt gcc agg gta  gct  5872 Tyr Asn Glu Met Val  Gly Glu Cys Trp Leu  # Gln Ser Ala Arg Val  Ala                 1920  #                1925  #                1930 aga aag gct ggt  cac cac cag aca gcc  t #ac aat gct ctc ctt  aat gca  5920 Arg Lys Ala Gly  His His Gln Thr Ala  T #yr Asn Ala Leu Leu  Asn Ala             1935      #            1940      #            1945 ggg gaa tca  cga ctc gct gaa ctg  tac g #tg gaa agg gca  aag tgg ctc  5968 Gly Glu Ser  Arg Leu Ala Glu Leu  Tyr V #al Glu Arg Ala  Lys Trp Leu         1950          #        1955          #        1960 tgg tcc  aag ggt gat gtt cac  cag gca c #ta att gtt  ctt caa aaa ggt  6016 Trp Ser  Lys Gly Asp Val His  Gln Ala L #eu Ile Val  Leu Gln Lys Gly     1965              #    1970              #    1975 gtt  gaa tta tgt ttt cct  gaa aat gaa a #cc cca  cct gag ggt aag aac  6064 Val  Glu Leu Cys Phe Pro  Glu Asn Glu T #hr Pro  Pro Glu Gly Lys Asn 1980                  #1985                  #1990                  #1995 atg tta atc cat ggt  cga gct atg cta cta  # gtg ggc cga ttt atg  gaa  6112 Met Leu Ile His Gly  Arg Ala Met Leu Leu  # Val Gly Arg Phe Met  Glu                 2000  #                2005  #                2010 gaa aca gct aac  ttt gaa agc aat gca  a #tt atg aaa aaa tat  aag gat  6160 Glu Thr Ala Asn  Phe Glu Ser Asn Ala  I #le Met Lys Lys Tyr  Lys Asp             2015      #            2020      #            2025 gtg acc gcg  tgc ctg cca gaa tgg  gag g #at ggg cat ttt  tac ctt gcc  6208 Val Thr Ala  Cys Leu Pro Glu Trp  Glu A #sp Gly His Phe  Tyr Leu Ala         2030          #        2035          #        2040 aag tac  tat gac aaa ttg atg  ccc atg g #tc aca gac  aac aaa atg gaa  6256 Lys Tyr  Tyr Asp Lys Leu Met  Pro Met V #al Thr Asp  Asn Lys Met Glu     2045              #    2050              #    2055 aag  caa ggt gat ctc atc  cgg tat ata g #tt ctt  cat ttt ggc aga tct  6304 Lys  Gln Gly Asp Leu Ile  Arg Tyr Ile V #al Leu  His Phe Gly Arg Ser 2060                  #2065                  #2070                  #2075 cta caa tat gga aat  cag ttc ata tat cag  # tca atg cca cga atg  tta  6352 Leu Gln Tyr Gly Asn  Gln Phe Ile Tyr Gln  # Ser Met Pro Arg Met  Leu                 2080  #                2085  #                2090 act cta tgg ctt  gat tat ggt aca aag  g #ca tat gaa tgg gaa  aaa gct  6400 Thr Leu Trp Leu  Asp Tyr Gly Thr Lys  A #la Tyr Glu Trp Glu  Lys Ala             2095      #            2100      #            2105 ggc cgc tcc  gat cgt gta caa atg  agg a #at gat ttg ggt  aaa ata aac  6448 Gly Arg Ser  Asp Arg Val Gln Met  Arg A #sn Asp Leu Gly  Lys Ile Asn         2110          #        2115          #        2120 aag gtt  atc aca gag cat aca  aac tat t #ta gct cca  tat caa ttt ttg  6496 Lys Val  Ile Thr Glu His Thr  Asn Tyr L #eu Ala Pro  Tyr Gln Phe Leu     2125              #    2130              #    2135 act  gct ttt tca caa ttg  atc tct cga a #tt tgt  cat tct cac gat gaa  6544 Thr  Ala Phe Ser Gln Leu  Ile Ser Arg I #le Cys  His Ser His Asp Glu 2140                  #2145                  #2150                  #2155 gtt ttt gtt gtc ttg  atg gaa ata ata gcc  # aaa gta ttt cta gcc  tat  6592 Val Phe Val Val Leu  Met Glu Ile Ile Ala  # Lys Val Phe Leu Ala  Tyr                 2160  #                2165  #                2170 cct caa caa gca  atg tgg atg atg aca  g #ct gtg tca aag tca  tct tat  6640 Pro Gln Gln Ala  Met Trp Met Met Thr  A #la Val Ser Lys Ser  Ser Tyr             2175      #            2180      #            2185 ccc atg cgt  gtg aac aga tgc aag  gaa a #tc ctc aat aaa  gct att cat  6688 Pro Met Arg  Val Asn Arg Cys Lys  Glu I #le Leu Asn Lys  Ala Ile His         2190          #        2195          #        2200 atg aaa  aaa tcc tta gag aag  ttt gtt g #ga gat gca  act cgc cta aca  6736 Met Lys  Lys Ser Leu Glu Lys  Phe Val G #ly Asp Ala  Thr Arg Leu Thr     2205              #    2210              #    2215 gat  aag ctt cta gaa ttg  tgc aat aaa c #cg gtt  gat gga agt agt tcc  6784 Asp  Lys Leu Leu Glu Leu  Cys Asn Lys P #ro Val  Asp Gly Ser Ser Ser 2220                  #2225                  #2230                  #2235 aca tta agc atg agc  act cat ttt aaa atg  # ctt aaa aag ctg gta  gaa  6832 Thr Leu Ser Met Ser  Thr His Phe Lys Met  # Leu Lys Lys Leu Val  Glu                 2240  #                2245  #                2250 gaa gca aca ttt  agt gaa atc ctc att  c #ct cta caa tca gtc  atg ata  6880 Glu Ala Thr Phe  Ser Glu Ile Leu Ile  P #ro Leu Gln Ser Val  Met Ile             2255      #            2260      #            2265 cct aca ctt  cca tca att ctg ggt  acc c #at gct aac cat  gct agc cat  6928 Pro Thr Leu  Pro Ser Ile Leu Gly  Thr H #is Ala Asn His  Ala Ser His         2270          #        2275          #        2280 gaa cca  ttt cct gga cat tgg  gcc tat a #tt gca ggg  ttt gat gat atg  6976 Glu Pro  Phe Pro Gly His Trp  Ala Tyr I #le Ala Gly  Phe Asp Asp Met     2285              #    2290              #    2295 gtg  gaa att ctt gct tct  ctt cag aaa c #ca aag  aag att tct tta aaa  7024 Val  Glu Ile Leu Ala Ser  Leu Gln Lys P #ro Lys  Lys Ile Ser Leu Lys 2300                  #2305                  #2310                  #2315 ggc tca gat gga aag  ttc tac atc atg atg  # tgt aag cca aaa gat gac   7072 Gly Ser Asp Gly Lys  Phe Tyr Ile Met Met  # Cys Lys Pro Lys Asp  Asp                 2320  #                2325  #                2330 ctg aga aag gat  tgt aga cta atg gaa  t #tc aat tcc ttg att  aat aag  7120 Leu Arg Lys Asp  Cys Arg Leu Met Glu  P #he Asn Ser Leu Ile  Asn Lys             2335      #            2340      #            2345 tgc tta aga  aaa gat gca gag tct  cgt a #ga aga gaa ctt  cat att cga  7168 Cys Leu Arg  Lys Asp Ala Glu Ser  Arg A #rg Arg Glu Leu  His Ile Arg         2350          #        2355          #        2360 aca tat  gca gtt att cca cta  aat gat g #aa tgt ggg  att att gaa tgg  7216 Thr Tyr  Ala Val Ile Pro Leu  Asn Asp G #lu Cys Gly  Ile Ile Glu Trp     2365              #    2370              #    2375 gtg  aac aac act gct ggt  ttg aga cct a #tt ctg  acc aaa cta tat aaa  7264 Val  Asn Asn Thr Ala Gly  Leu Arg Pro I #le Leu  Thr Lys Leu Tyr Lys 2380                  #2385                  #2390                  #2395 gaa aag gga gtg tat  atg aca gga aaa gaa  # ctt cgc cag tgt atg  cta  7312 Glu Lys Gly Val Tyr  Met Thr Gly Lys Glu  # Leu Arg Gln Cys Met  Leu                 2400  #                2405  #                2410 cca aag tca gca  gct tta tct gaa aaa  c #tc aaa gta ttc cga  gaa ttt  7360 Pro Lys Ser Ala  Ala Leu Ser Glu Lys  L #eu Lys Val Phe Arg  Glu Phe             2415      #            2420      #            2425 ctc ctg ccc  agg cat cct cct att  ttt c #at gag tgg ttt  ctg aga aca  7408 Leu Leu Pro  Arg His Pro Pro Ile  Phe H #is Glu Trp Phe  Leu Arg Thr         2430          #        2435          #        2440 ttc cct  gat cct aca tca tgg  tac agt a #gt aga tca  gct tac tgc cgt  7456 Phe Pro  Asp Pro Thr Ser Trp  Tyr Ser S #er Arg Ser  Ala Tyr Cys Arg     2445              #    2450              #    2455 tcc  act gca gta atg tca  atg gtt ggt t #at att  ctg ggg ctt gga gac  7504 Ser  Thr Ala Val Met Ser  Met Val Gly T #yr Ile  Leu Gly Leu Gly Asp 2460                  #2465                  #2470                  #2475 cgt cat ggt gaa aat  att ctc ttt gat tct  # ttg act ggt gaa tgc  gta  7552 Arg His Gly Glu Asn  Ile Leu Phe Asp Ser  # Leu Thr Gly Glu Cys  Val                 2480  #                2485  #                2490 cat gta gat ttc  aat tgt ctt ttc aat  a #ag gga gaa acc ttt  gaa gtt  7600 His Val Asp Phe  Asn Cys Leu Phe Asn  L #ys Gly Glu Thr Phe  Glu Val             2495      #            2500      #            2505 cca gaa att  gtg cca ttt cgc ctg  act c #at aat atg gtt  aat gga atg  7648 Pro Glu Ile  Val Pro Phe Arg Leu  Thr H #is Asn Met Val  Asn Gly Met         2510          #        2515          #        2520 ggt cct  atg gga aca gag ggt  ctt ttt c #ga aga gca  tgt gaa gtt aca  7696 Gly Pro  Met Gly Thr Glu Gly  Leu Phe A #rg Arg Ala  Cys Glu Val Thr     2525              #    2530              #    2535 atg  agg ctg atg cgt gat  cag cga gag c #ct tta  atg agt gtc tta aag  7744 Met  Arg Leu Met Arg Asp  Gln Arg Glu P #ro Leu  Met Ser Val Leu Lys 2540                  #2545                  #2550                  #2555 act ttt cta cat gat  cct ctt gtg gaa tgg  # agt aaa cca gtg aaa  ggg  7792 Thr Phe Leu His Asp  Pro Leu Val Glu Trp  # Ser Lys Pro Val Lys  Gly                 2560  #                2565  #                2570 cat tcc aaa gcg  cca ctg aat gaa act  g #ga gaa gtt gtc aat  gaa aag  7840 His Ser Lys Ala  Pro Leu Asn Glu Thr  G #ly Glu Val Val Asn  Glu Lys             2575      #            2580      #            2585 gcc aag acc  cat gtt ctt gac att  gag c #ag cga cta caa  ggt gta atc  7888 Ala Lys Thr  His Val Leu Asp Ile  Glu G #ln Arg Leu Gln  Gly Val Ile         2590          #        2595          #        2600 aag act  cga aat aga gtg aca  gga ctg c #cg tta tct  att gaa gga cat  7936 Lys Thr  Arg Asn Arg Val Thr  Gly Leu P #ro Leu Ser  Ile Glu Gly His     2605              #    2610              #    2615 gtg  cat tac ctt ata caa  gaa gct act g #at gaa  aac tta cta tgc cag  7984 Val  His Tyr Leu Ile Gln  Glu Ala Thr A #sp Glu  Asn Leu Leu Cys Gln 2620                  #2625                  #2630                  #2635 atg tat ctt ggt tgg  act cca tat atg tgaaa #tgaaa ttatgtaaaa          8031 Met Tyr Leu Gly Trp  Thr Pro Tyr Met                 2640 gaatatgtta ataatctaaa agtaatgcat ttggtatgaa tctgtggttg ta #tctgttca   8091 attctaaagt acaacataaa tttacgttct cagcaactgt tatttctctc tg #atcattaa   8151 ttatatgtaa aataatatac attcagttat taagaaataa actgctttct ta #ataaaaaa   8211 aaaaaaaaaa aaaaaaaaaa aaaaaaaa          #                   #           8239 <210> SEQ ID NO 2 <211> LENGTH: 2644 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 2 Met Gly Glu His Gly Leu Glu Leu Ala Ser Me #t Ile Pro Ala Leu Arg 1               5    #                10   #                15 Glu Leu Gly Ser Ala Thr Pro Glu Glu Tyr As #n Thr Val Val Gln Lys             20       #            25       #            30 Pro Arg Gln Ile Leu Cys Gln Phe Ile Asp Ar #g Ile Leu Thr Asp Val         35           #        40           #        45 Asn Val Val Ala Val Glu Leu Val Lys Lys Th #r Asp Ser Gln Pro Thr     50               #    55               #    60 Ser Val Met Leu Leu Asp Phe Ile Gln His Il #e Met Lys Ser Ser Pro 65                   #70                   #75                   #80 Leu Met Phe Val Asn Val Ser Gly Ser His Gl #u Arg Lys Gly Ser Cys                 85   #                90   #                95 Ile Glu Phe Ser Asn Trp Ile Ile Thr Arg Le #u Leu Arg Ile Ala Ala             100       #           105       #           110 Thr Pro Ser Cys His Leu Leu His Lys Lys Il #e Cys Glu Val Ile Cys         115           #       120           #       125 Ser Leu Leu Phe Leu Phe Lys Ser Lys Ser Pr #o Ala Ile Phe Gly Val     130               #   135               #   140 Leu Thr Lys Glu Leu Leu Gln Leu Phe Glu As #p Leu Val Tyr Leu His 145                 1 #50                 1 #55                 1 #60 Arg Arg Asn Val Met Gly His Ala Val Glu Tr #p Pro Val Val Met Ser                 165   #               170   #               175 Arg Phe Leu Ser Gln Leu Asp Glu His Met Gl #y Tyr Leu Gln Ser Ala             180       #           185       #           190 Pro Leu Gln Leu Met Ser Met Gln Asn Leu Gl #u Phe Ile Glu Val Thr         195           #       200           #       205 Leu Leu Met Val Leu Thr Arg Ile Ile Ala Il #e Val Phe Phe Arg Arg     210               #   215               #   220 Gln Glu Leu Leu Leu Trp Gln Ile Gly Cys Va #l Leu Leu Glu Tyr Gly 225                 2 #30                 2 #35                 2 #40 Ser Pro Lys Ile Lys Ser Leu Ala Ile Ser Ph #e Leu Thr Glu Leu Phe                 245   #               250   #               255 Gln Leu Gly Gly Leu Pro Ala Gln Pro Ala Se #r Thr Phe Phe Ser Ser             260       #           265       #           270 Phe Leu Glu Leu Leu Lys His Leu Val Glu Me #t Asp Thr Asp Gln Leu         275           #       280           #       285 Lys Leu Tyr Glu Glu Pro Leu Ser Lys Leu Il #e Lys Thr Leu Phe Pro     290               #   295               #   300 Phe Glu Ala Glu Ala Tyr Arg Asn Ile Glu Pr #o Val Tyr Leu Asn Met 305                 3 #10                 3 #15                 3 #20 Leu Leu Glu Lys Leu Cys Val Met Phe Glu As #p Gly Val Leu Met Arg                 325   #               330   #               335 Leu Lys Ser Asp Leu Leu Lys Ala Ala Leu Cy #s His Leu Leu Gln Tyr             340       #           345       #           350 Phe Leu Lys Phe Val Pro Ala Gly Tyr Glu Se #r Ala Leu Gln Val Arg         355           #       360           #       365 Lys Val Tyr Val Arg Asn Ile Cys Lys Ala Le #u Leu Asp Val Leu Gly     370               #   375               #   380 Ile Glu Val Asp Ala Glu Tyr Leu Leu Gly Pr #o Leu Tyr Ala Ala Leu 385                 3 #90                 3 #95                 4 #00 Lys Met Glu Ser Met Glu Ile Ile Glu Glu Il #e Gln Cys Gln Thr Gln                 405   #               410   #               415 Gln Glu Asn Leu Ser Ser Asn Ser Asp Gly Il #e Ser Pro Lys Arg Arg             420       #           425       #           430 Arg Leu Ser Ser Ser Leu Asn Pro Ser Lys Ar #g Ala Pro Lys Gln Thr         435           #       440           #       445 Glu Glu Ile Lys His Val Asp Met Asn Gln Ly #s Ser Ile Leu Trp Ser     450               #   455               #   460 Ala Leu Lys Gln Lys Ala Glu Ser Leu Gln Il #e Ser Leu Glu Tyr Ser 465                 4 #70                 4 #75                 4 #80 Gly Leu Lys Asn Pro Val Ile Glu Met Leu Gl #u Gly Ile Ala Val Val                 485   #               490   #               495 Leu Gln Leu Thr Ala Leu Cys Thr Val His Cy #s Ser His Gln Asn Met             500       #           505       #           510 Asn Cys Arg Thr Phe Lys Asp Cys Gln His Ly #s Ser Lys Lys Lys Pro         515           #       520           #       525 Ser Val Val Ile Thr Trp Met Ser Leu Asp Ph #e Tyr Thr Lys Val Leu     530               #   535               #   540 Lys Ser Cys Arg Ser Leu Leu Glu Ser Val Gl #n Lys Leu Asp Leu Glu 545                 5 #50                 5 #55                 5 #60 Ala Thr Ile Asp Lys Val Val Lys Ile Tyr As #p Ala Leu Ile Tyr Met                 565   #               570   #               575 Gln Val Asn Ser Ser Phe Glu Asp His Ile Le #u Glu Asp Leu Cys Gly             580       #           585       #           590 Met Leu Ser Leu Pro Trp Ile Tyr Ser His Se #r Asp Asp Gly Cys Leu         595           #       600           #       605 Lys Leu Thr Thr Phe Ala Ala Asn Leu Leu Th #r Leu Ser Cys Arg Ile     610               #   615               #   620 Ser Asp Ser Tyr Ser Pro Gln Ala Gln Ser Ar #g Cys Val Phe Leu Leu 625                 6 #30                 6 #35                 6 #40 Thr Leu Phe Pro Arg Arg Ile Phe Leu Glu Tr #p Arg Thr Ala Val Tyr                 645   #               650   #               655 Asn Trp Ala Leu Gln Ser Ser His Glu Val Il #e Arg Ala Ser Cys Val             660       #           665       #           670 Ser Gly Phe Phe Ile Leu Leu Gln Gln Gln As #n Ser Cys Asn Arg Val         675           #       680           #       685 Pro Lys Ile Leu Ile Asp Lys Val Lys Asp As #p Ser Asp Ile Val Lys     690               #   695               #   700 Lys Glu Phe Ala Ser Ile Leu Gly Gln Leu Va #l Cys Thr Leu His Gly 705                 7 #10                 7 #15                 7 #20 Met Phe Tyr Leu Thr Ser Ser Leu Thr Glu Pr #o Phe Ser Glu His Gly                 725   #               730   #               735 His Val Asp Leu Phe Cys Arg Asn Leu Lys Al #a Thr Ser Gln His Glu             740       #           745       #           750 Cys Ser Ser Ser Gln Leu Lys Ala Ser Val Cy #s Lys Pro Phe Leu Phe         755           #       760           #       765 Leu Leu Lys Lys Lys Ile Pro Ser Pro Val Ly #s Leu Ala Phe Ile Asp     770               #   775               #   780 Asn Leu His His Leu Cys Lys His Leu Asp Ph #e Arg Glu Asp Glu Thr 785                 7 #90                 7 #95                 8 #00 Asp Val Lys Ala Val Leu Gly Thr Leu Leu As #n Leu Met Glu Asp Pro                 805   #               810   #               815 Asp Lys Asp Val Arg Val Ala Phe Ser Gly As #n Ile Lys His Ile Leu             820       #           825       #           830 Glu Ser Leu Asp Ser Glu Asp Gly Phe Ile Ly #s Glu Leu Phe Val Leu         835           #       840           #       845 Arg Met Lys Glu Ala Tyr Thr His Ala Gln Il #e Ser Arg Asn Asn Glu     850               #   855               #   860 Leu Lys Asp Thr Leu Ile Leu Thr Thr Gly As #p Ile Gly Arg Ala Ala 865                 8 #70                 8 #75                 8 #80 Lys Gly Asp Leu Val Pro Phe Ala Leu Leu Hi #s Leu Leu His Cys Leu                 885   #               890   #               895 Leu Ser Lys Ser Ala Ser Val Ser Gly Ala Al #a Tyr Thr Glu Ile Arg             900       #           905       #           910 Ala Leu Val Ala Ala Lys Ser Val Lys Leu Gl #n Ser Phe Phe Ser Gln         915           #       920           #       925 Tyr Lys Lys Pro Ile Cys Gln Phe Leu Val Gl #u Ser Leu His Ser Ser     930               #   935               #   940 Gln Met Thr Ala Leu Pro Asn Thr Pro Cys Gl #n Asn Ala Asp Val Arg 945                 9 #50                 9 #55                 9 #60 Lys Gln Asp Val Ala His Gln Arg Glu Met Al #a Leu Asn Thr Leu Ser                 965   #               970   #               975 Glu Ile Ala Asn Val Phe Asp Phe Pro Asp Le #u Asn Arg Phe Leu Thr             980       #           985       #           990 Arg Thr Leu Gln Val Leu Leu Pro  Asp Leu  #Ala Ala Lys  Ala Ser Pro         995           #       1000           #       1005 Ala Ala  Ser Ala Leu Ile Arg  Thr Leu G #ly Lys Gln  Leu Asn Val     1010              #    1015              #    1020 Asn Arg  Arg Glu Ile Leu Ile  Asn Asn P #he Lys Tyr  Ile Phe Ser     1025              #    1030              #    1035 His Leu  Val Cys Ser Cys Ser  Lys Asp G #lu Leu Glu  Arg Ala Leu     1040              #    1045              #    1050 His Tyr  Leu Lys Asn Glu Thr  Glu Ile G #lu Leu Gly  Ser Leu Leu     1055              #    1060              #    1065 Arg Gln  Asp Phe Gln Gly Leu  His Asn G #lu Leu Leu  Leu Arg Ile     1070              #    1075              #    1080 Gly Glu  His Tyr Gln Gln Val  Phe Asn G #ly Leu Ser  Ile Leu Ala     1085              #    1090              #    1095 Ser Phe  Ala Ser Ser Asp Asp  Pro Tyr G #ln Gly Pro  Arg Asp Ile     1100              #    1105              #    1110 Ile Ser  Pro Glu Leu Met Ala  Asp Tyr L #eu Gln Pro  Lys Leu Leu     1115              #    1120              #    1125 Gly Ile  Leu Ala Phe Phe Asn  Met Gln L #eu Leu Ser  Ser Ser Val     1130              #    1135              #    1140 Gly Ile  Glu Asp Lys Lys Met  Ala Leu A #sn Ser Leu  Met Ser Leu     1145              #    1150              #    1155 Met Lys  Leu Met Gly Pro Lys  His Val S #er Ser Val  Arg Val Lys     1160              #    1165              #    1170 Met Met  Thr Thr Leu Arg Thr  Gly Leu A #rg Phe Lys  Asp Asp Phe     1175              #    1180              #    1185 Pro Glu  Leu Cys Cys Arg Ala  Trp Asp C #ys Phe Val  Arg Cys Leu     1190              #    1195              #    1200 Asp His  Ala Cys Leu Gly Ser  Leu Leu S #er His Val  Ile Val Ala     1205              #    1210              #    1215 Leu Leu  Pro Leu Ile His Ile  Gln Pro L #ys Glu Thr  Ala Ala Ile     1220              #    1225              #    1230 Phe His  Tyr Leu Ile Ile Glu  Asn Arg A #sp Ala Val  Gln Asp Phe     1235              #    1240              #    1245 Leu His  Glu Ile Tyr Phe Leu  Pro Asp H #is Pro Glu  Leu Lys Lys     1250              #    1255              #    1260 Ile Lys  Ala Val Leu Gln Glu  Tyr Arg L #ys Glu Thr  Ser Glu Ser     1265              #    1270              #    1275 Thr Asp  Leu Gln Thr Thr Leu  Gln Leu S #er Met Lys  Ala Ile Gln     1280              #    1285              #    1290 His Glu  Asn Val Asp Val Arg  Ile His A #la Leu Thr  Ser Leu Lys     1295              #    1300              #    1305 Glu Thr  Leu Tyr Lys Asn Gln  Glu Lys L #eu Ile Lys  Tyr Ala Thr     1310              #    1315              #    1320 Asp Ser  Glu Thr Val Glu Pro  Ile Ile S #er Gln Leu  Val Thr Val     1325              #    1330              #    1335 Leu Leu  Lys Gly Cys Gln Asp  Ala Asn S #er Gln Ala  Arg Leu Leu     1340              #    1345              #    1350 Cys Gly  Glu Cys Leu Gly Glu  Leu Gly A #la Ile Asp  Pro Gly Arg     1355              #    1360              #    1365 Leu Asp  Phe Ser Thr Thr Glu  Thr Gln G #ly Lys Asp  Phe Thr Phe     1370              #    1375              #    1380 Val Thr  Gly Val Glu Asp Ser  Ser Phe A #la Tyr Gly  Leu Leu Met     1385              #    1390              #    1395 Glu Leu  Thr Arg Ala Tyr Leu  Ala Tyr A #la Asp Asn  Ser Arg Ala     1400              #    1405              #    1410 Gln Asp  Ser Ala Ala Tyr Ala  Ile Gln G #lu Leu Leu  Ser Ile Tyr     1415              #    1420              #    1425 Asp Cys  Arg Glu Met Glu Thr  Asn Gly P #ro Gly His  Gln Leu Trp     1430              #    1435              #    1440 Arg Arg  Phe Pro Glu His Val  Arg Glu I #le Leu Glu  Pro His Leu     1445              #    1450              #    1455 Asn Thr  Arg Tyr Lys Ser Ser  Gln Lys S #er Thr Asp  Trp Ser Gly     1460              #    1465              #    1470 Val Lys  Lys Pro Ile Tyr Leu  Ser Lys L #eu Gly Ser  Asn Phe Ala     1475              #    1480              #    1485 Glu Trp  Ser Ala Ser Trp Ala  Gly Tyr L #eu Ile Thr  Lys Val Arg     1490              #    1495              #    1500 His Asp  Leu Ala Ser Lys Ile  Phe Thr C #ys Cys Ser  Ile Met Met     1505              #    1510              #    1515 Lys His  Asp Phe Lys Val Thr  Ile Tyr L #eu Leu Pro  His Ile Leu     1520              #    1525              #    1530 Val Tyr  Val Leu Leu Gly Cys  Asn Gln G #lu Asp Gln  Gln Glu Val     1535              #    1540              #    1545 Tyr Ala  Glu Ile Met Ala Val  Leu Lys H #is Asp Asp  Gln His Thr     1550              #    1555              #    1560 Ile Asn  Thr Gln Asp Ile Ala  Ser Asp L #eu Cys Gln  Leu Ser Thr     1565              #    1570              #    1575 Gln Thr  Val Phe Ser Met Leu  Asp His L #eu Thr Gln  Trp Ala Arg     1580              #    1585              #    1590 His Lys  Phe Gln Ala Leu Lys  Ala Glu L #ys Cys Pro  His Ser Lys     1595              #    1600              #    1605 Ser Asn  Arg Asn Lys Val Asp  Ser Met V #al Ser Thr  Val Asp Tyr     1610              #    1615              #    1620 Glu Asp  Tyr Gln Ser Val Thr  Arg Phe L #eu Asp Leu  Ile Pro Gln     1625              #    1630              #    1635 Asp Thr  Leu Ala Val Ala Ser  Phe Arg S #er Lys Ala  Tyr Thr Arg     1640              #    1645              #    1650 Ala Val  Met His Phe Glu Ser  Phe Ile T #hr Glu Lys  Lys Gln Asn     1655              #    1660              #    1665 Ile Gln  Glu His Leu Gly Phe  Leu Gln L #ys Leu Tyr  Ala Ala Met     1670              #    1675              #    1680 His Glu  Pro Asp Gly Val Ala  Gly Val S #er Ala Ile  Arg Lys Ala     1685              #    1690              #    1695 Glu Pro  Ser Leu Lys Glu Gln  Ile Leu G #lu His Glu  Ser Leu Gly     1700              #    1705              #    1710 Leu Leu  Arg Asp Ala Thr Ala  Cys Tyr A #sp Arg Ala  Ile Gln Leu     1715              #    1720              #    1725 Glu Pro  Asp Gln Ile Ile His  Tyr His G #ly Val Val  Lys Ser Met     1730              #    1735              #    1740 Leu Gly  Leu Gly Gln Leu Ser  Thr Val I #le Thr Gln  Val Asn Gly     1745              #    1750              #    1755 Val His  Ala Asn Arg Ser Glu  Trp Thr A #sp Glu Leu  Asn Thr Tyr     1760              #    1765              #    1770 Arg Val  Glu Ala Ala Trp Lys  Leu Ser G #ln Trp Asp  Leu Val Glu     1775              #    1780              #    1785 Asn Tyr  Leu Ala Ala Asp Gly  Lys Ser T #hr Thr Trp  Ser Val Arg     1790              #    1795              #    1800 Leu Gly  Gln Leu Leu Leu Ser  Ala Lys L #ys Arg Asp  Ile Thr Ala     1805              #    1810              #    1815 Phe Tyr  Asp Ser Leu Lys Leu  Val Arg A #la Glu Gln  Ile Val Pro     1820              #    1825              #    1830 Leu Ser  Ala Ala Ser Phe Glu  Arg Gly S #er Tyr Gln  Arg Gly Tyr     1835              #    1840              #    1845 Glu Tyr  Ile Val Arg Leu His  Met Leu C #ys Glu Leu  Glu His Ser     1850              #    1855              #    1860 Ile Lys  Pro Leu Phe Gln His  Ser Pro G #ly Asp Ser  Ser Gln Glu     1865              #    1870              #    1875 Asp Ser  Leu Asn Trp Val Ala  Arg Leu G #lu Met Thr  Gln Asn Ser     1880              #    1885              #    1890 Tyr Arg  Ala Lys Asp Pro Ile  Leu Ala L #eu Arg Arg  Ala Leu Leu     1895              #    1900              #    1905 Ser Leu  Asn Lys Arg Pro Asp  Tyr Asn G #lu Met Val  Gly Glu Cys     1910              #    1915              #    1920 Trp Leu  Gln Ser Ala Arg Val  Ala Arg L #ys Ala Gly  His His Gln     1925              #    1930              #    1935 Thr Ala  Tyr Asn Ala Leu Leu  Asn Ala G #ly Glu Ser  Arg Leu Ala     1940              #    1945              #    1950 Glu Leu  Tyr Val Glu Arg Ala  Lys Trp L #eu Trp Ser  Lys Gly Asp     1955              #    1960              #    1965 Val His  Gln Ala Leu Ile Val  Leu Gln L #ys Gly Val  Glu Leu Cys     1970              #    1975              #    1980 Phe Pro  Glu Asn Glu Thr Pro  Pro Glu G #ly Lys Asn  Met Leu Ile     1985              #    1990              #    1995 His Gly  Arg Ala Met Leu Leu  Val Gly A #rg Phe Met  Glu Glu Thr     2000              #    2005              #    2010 Ala Asn  Phe Glu Ser Asn Ala  Ile Met L #ys Lys Tyr  Lys Asp Val     2015              #    2020              #    2025 Thr Ala  Cys Leu Pro Glu Trp  Glu Asp G #ly His Phe  Tyr Leu Ala     2030              #    2035              #    2040 Lys Tyr  Tyr Asp Lys Leu Met  Pro Met V #al Thr Asp  Asn Lys Met     2045              #    2050              #    2055 Glu Lys  Gln Gly Asp Leu Ile  Arg Tyr I #le Val Leu  His Phe Gly     2060              #    2065              #    2070 Arg Ser  Leu Gln Tyr Gly Asn  Gln Phe I #le Tyr Gln  Ser Met Pro     2075              #    2080              #    2085 Arg Met  Leu Thr Leu Trp Leu  Asp Tyr G #ly Thr Lys  Ala Tyr Glu     2090              #    2095              #    2100 Trp Glu  Lys Ala Gly Arg Ser  Asp Arg V #al Gln Met  Arg Asn Asp     2105              #    2110              #    2115 Leu Gly  Lys Ile Asn Lys Val  Ile Thr G #lu His Thr  Asn Tyr Leu     2120              #    2125              #    2130 Ala Pro  Tyr Gln Phe Leu Thr  Ala Phe S #er Gln Leu  Ile Ser Arg     2135              #    2140              #    2145 Ile Cys  His Ser His Asp Glu  Val Phe V #al Val Leu  Met Glu Ile     2150              #    2155              #    2160 Ile Ala  Lys Val Phe Leu Ala  Tyr Pro G #ln Gln Ala  Met Trp Met     2165              #    2170              #    2175 Met Thr  Ala Val Ser Lys Ser  Ser Tyr P #ro Met Arg  Val Asn Arg     2180              #    2185              #    2190 Cys Lys  Glu Ile Leu Asn Lys  Ala Ile H #is Met Lys  Lys Ser Leu     2195              #    2200              #    2205 Glu Lys  Phe Val Gly Asp Ala  Thr Arg L #eu Thr Asp  Lys Leu Leu     2210              #    2215              #    2220 Glu Leu  Cys Asn Lys Pro Val  Asp Gly S #er Ser Ser  Thr Leu Ser     2225              #    2230              #    2235 Met Ser  Thr His Phe Lys Met  Leu Lys L #ys Leu Val  Glu Glu Ala     2240              #    2245              #    2250 Thr Phe  Ser Glu Ile Leu Ile  Pro Leu G #ln Ser Val  Met Ile Pro     2255              #    2260              #    2265 Thr Leu  Pro Ser Ile Leu Gly  Thr His A #la Asn His  Ala Ser His     2270              #    2275              #    2280 Glu Pro  Phe Pro Gly His Trp  Ala Tyr I #le Ala Gly  Phe Asp Asp     2285              #    2290              #    2295 Met Val  Glu Ile Leu Ala Ser  Leu Gln L #ys Pro Lys  Lys Ile Ser     2300              #    2305              #    2310 Leu Lys  Gly Ser Asp Gly Lys  Phe Tyr I #le Met Met  Cys Lys Pro     2315              #    2320              #    2325 Lys Asp  Asp Leu Arg Lys Asp  Cys Arg L #eu Met Glu  Phe Asn Ser     2330              #    2335              #    2340 Leu Ile  Asn Lys Cys Leu Arg  Lys Asp A #la Glu Ser  Arg Arg Arg     2345              #    2350              #    2355 Glu Leu  His Ile Arg Thr Tyr  Ala Val I #le Pro Leu  Asn Asp Glu     2360              #    2365              #    2370 Cys Gly  Ile Ile Glu Trp Val  Asn Asn T #hr Ala Gly  Leu Arg Pro     2375              #    2380              #    2385 Ile Leu  Thr Lys Leu Tyr Lys  Glu Lys G #ly Val Tyr  Met Thr Gly     2390              #    2395              #    2400 Lys Glu  Leu Arg Gln Cys Met  Leu Pro L #ys Ser Ala  Ala Leu Ser     2405              #    2410              #    2415 Glu Lys  Leu Lys Val Phe Arg  Glu Phe L #eu Leu Pro  Arg His Pro     2420              #    2425              #    2430 Pro Ile  Phe His Glu Trp Phe  Leu Arg T #hr Phe Pro  Asp Pro Thr     2435              #    2440              #    2445 Ser Trp  Tyr Ser Ser Arg Ser  Ala Tyr C #ys Arg Ser  Thr Ala Val     2450              #    2455              #    2460 Met Ser  Met Val Gly Tyr Ile  Leu Gly L #eu Gly Asp  Arg His Gly     2465              #    2470              #    2475 Glu Asn  Ile Leu Phe Asp Ser  Leu Thr G #ly Glu Cys  Val His Val     2480              #    2485              #    2490 Asp Phe  Asn Cys Leu Phe Asn  Lys Gly G #lu Thr Phe  Glu Val Pro     2495              #    2500              #    2505 Glu Ile  Val Pro Phe Arg Leu  Thr His A #sn Met Val  Asn Gly Met     2510              #    2515              #    2520 Gly Pro  Met Gly Thr Glu Gly  Leu Phe A #rg Arg Ala  Cys Glu Val     2525              #    2530              #    2535 Thr Met  Arg Leu Met Arg Asp  Gln Arg G #lu Pro Leu  Met Ser Val     2540              #    2545              #    2550 Leu Lys  Thr Phe Leu His Asp  Pro Leu V #al Glu Trp  Ser Lys Pro     2555              #    2560              #    2565 Val Lys  Gly His Ser Lys Ala  Pro Leu A #sn Glu Thr  Gly Glu Val     2570              #    2575              #    2580 Val Asn  Glu Lys Ala Lys Thr  His Val L #eu Asp Ile  Glu Gln Arg     2585              #    2590              #    2595 Leu Gln  Gly Val Ile Lys Thr  Arg Asn A #rg Val Thr  Gly Leu Pro     2600              #    2605              #    2610 Leu Ser  Ile Glu Gly His Val  His Tyr L #eu Ile Gln  Glu Ala Thr     2615              #    2620              #    2625 Asp Glu  Asn Leu Leu Cys Gln  Met Tyr L #eu Gly Trp  Thr Pro Tyr     2630              #    2635              #    2640 Met <210> SEQ ID NO 3 <211> LENGTH: 8022 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: CDS <222> LOCATION: (585)..(7742) <223> OTHER INFORMATION: <400> SEQUENCE: 3 ggtaccaagt aaaaactgct tagtaagtat aaaacacaga agaatccgcg at #ctagtgaa     60 ccaatgccct gcgtatgacg ctccactgac gctatagtca atgagaacta gg #atgtgcga    120 ttataactta tcttttcaat attttcttat tatttattta agaaataatt ga #attaaaac    180 tcatttcttc ttttattagc cgtaaaatag cttattttct ctcctactac ct #ttcaacaa    240 taactttttt ttttgtttat tgaccattat aatcacatca aaagtcaaaa aa #ttcaatca    300 ttatcagaaa catccagcct aatattactt aaaagttagt ttcctctgaa aa #ttcagtat    360 cacaaaagct cgttaattag catcgctcga tacttagtgc accatgcatc tt #cctttacc    420 tcgtgagtgg aaatcgattt gataatcgat tgccactttt cgcataattc ta #ttgagata    480 ttttattact tacaatcgtc ttttataaat gctcaagact ttgaacgcgc gt #gttgcgtt    540 ttaaaaaggc ctttttttga attgaatcaa tggtttgata tagt atg a #gc caa cac     596                    #                   #             Met Ser Gln  #His                    #                   #             1 gca aaa agg aaa gct ggg tca ctc gat ctt tc #a ccc aga ggc tta gat      644 Ala Lys Arg Lys Ala Gly Ser Leu Asp Leu Se #r Pro Arg Gly Leu Asp 5                   # 10                  # 15                  # 20 gac aga cag gct ttc gga cag ctt ttg aaa ga #a gta tta gca tta gac      692 Asp Arg Gln Ala Phe Gly Gln Leu Leu Lys Gl #u Val Leu Ala Leu Asp                 25   #                30   #                35 aaa gaa cat gag tta ggt aga agt aat tct tt #a cca tct atg acc tcc      740 Lys Glu His Glu Leu Gly Arg Ser Asn Ser Le #u Pro Ser Met Thr Ser             40       #            45       #            50 gag ctt gtt gaa gtt tta att gaa gtt ggt ct #t cta gct ttt aaa cat      788 Glu Leu Val Glu Val Leu Ile Glu Val Gly Le #u Leu Ala Phe Lys His         55           #        60           #        65 gat gat tca aaa tct gaa ttt atc tct cct aa #g atg cta aaa gaa gcc      836 Asp Asp Ser Lys Ser Glu Phe Ile Ser Pro Ly #s Met Leu Lys Glu Ala     70               #    75               #    80 cat ctc tct cta caa gcg tta atg cta atc tt #a aaa agg tct ccg aca      884 His Leu Ser Leu Gln Ala Leu Met Leu Ile Le #u Lys Arg Ser Pro Thr 85                   #90                   #95                   #100 gtt ttg cgg gag att aaa tca tct gtt act ct #t ttg gat tgg att tta      932 Val Leu Arg Glu Ile Lys Ser Ser Val Thr Le #u Leu Asp Trp Ile Leu                 105   #               110   #               115 ccc agg act ata tca ttg ttt gct gat att cg #t ttt att aag tta ttt      980 Pro Arg Thr Ile Ser Leu Phe Ala Asp Ile Ar #g Phe Ile Lys Leu Phe             120       #           125       #           130 gac tca tta aaa gag ttt cat aag cta att ta #t cag cta atc agt gaa     1028 Asp Ser Leu Lys Glu Phe His Lys Leu Ile Ty #r Gln Leu Ile Ser Glu         135           #       140           #       145 aag tca ttc cta tgg gac tta tat gct tcg tt #t atg cgt tat tgg aaa     1076 Lys Ser Phe Leu Trp Asp Leu Tyr Ala Ser Ph #e Met Arg Tyr Trp Lys     150               #   155               #   160 tat tat att aca aac gtt tct tct ata gtt ct #c caa atc act aat gct     1124 Tyr Tyr Ile Thr Asn Val Ser Ser Ile Val Le #u Gln Ile Thr Asn Ala 165                 1 #70                 1 #75                 1 #80 aca ttc cct tac aag atg ccc tca ccc aat tc #t caa cca ttg cag agt     1172 Thr Phe Pro Tyr Lys Met Pro Ser Pro Asn Se #r Gln Pro Leu Gln Ser                 185   #               190   #               195 atc tcc cca aat tat cca acc cat cga gag ga #c aaa ttt gat tta ctt     1220 Ile Ser Pro Asn Tyr Pro Thr His Arg Glu As #p Lys Phe Asp Leu Leu             200       #           205       #           210 atc att aat ata gag gag gct tgt aca ttt tt #c ttt gaa agt gcc cat     1268 Ile Ile Asn Ile Glu Glu Ala Cys Thr Phe Ph #e Phe Glu Ser Ala His         215           #       220           #       225 ttt ttt gca caa tgc tca tat tta aag aaa tc #c aat ttt cct agt cca     1316 Phe Phe Ala Gln Cys Ser Tyr Leu Lys Lys Se #r Asn Phe Pro Ser Pro     230               #   235               #   240 cct ctc ttt aca gcg tgg act tgg atc aag cc #a tgt ttt ttt aat ttt     1364 Pro Leu Phe Thr Ala Trp Thr Trp Ile Lys Pr #o Cys Phe Phe Asn Phe 245                 2 #50                 2 #55                 2 #60 gtt att tta tta aaa cga atc agc atc gga ga #c tca cag ctc ttt cta     1412 Val Ile Leu Leu Lys Arg Ile Ser Ile Gly As #p Ser Gln Leu Phe Leu                 265   #               270   #               275 cat ttg cat tca cgt ata gtc caa act tta tg #c tgt ttt tcc ttg aat     1460 His Leu His Ser Arg Ile Val Gln Thr Leu Cy #s Cys Phe Ser Leu Asn             280       #           285       #           290 ttt ata tat cat ggc ctt ccc att tgt gaa aa #a tct aaa cat att tta     1508 Phe Ile Tyr His Gly Leu Pro Ile Cys Glu Ly #s Ser Lys His Ile Leu         295           #       300           #       305 atg tcc tcc atc aac tta aca ttg gga tca tt #g aag aaa act tat aca     1556 Met Ser Ser Ile Asn Leu Thr Leu Gly Ser Le #u Lys Lys Thr Tyr Thr     310               #   315               #   320 gtt gct aat act gct ata tct ctt ttt ttt ct #c tct tta ttt gtt tta     1604 Val Ala Asn Thr Ala Ile Ser Leu Phe Phe Le #u Ser Leu Phe Val Leu 325                 3 #30                 3 #35                 3 #40 ccc aaa act gta gct ggt cta ttc tat cct tt #t ggg gtt tcc tta ctt     1652 Pro Lys Thr Val Ala Gly Leu Phe Tyr Pro Ph #e Gly Val Ser Leu Leu                 345   #               350   #               355 tct gac ttc aag gta tta gag caa ctt gaa cc #a gat tct gat ctc aaa     1700 Ser Asp Phe Lys Val Leu Glu Gln Leu Glu Pr #o Asp Ser Asp Leu Lys             360       #           365       #           370 aag gca ata ata tta ttt aag tgc aga tac ca #a agt tca gaa ata gat     1748 Lys Ala Ile Ile Leu Phe Lys Cys Arg Tyr Gl #n Ser Ser Glu Ile Asp         375           #       380           #       385 caa aca act ctc cgt gct ttt ggc gaa att tg #t act ggt aaa ctt gaa     1796 Gln Thr Thr Leu Arg Ala Phe Gly Glu Ile Cy #s Thr Gly Lys Leu Glu     390               #   395               #   400 aac acg ttg ttt tct aac tct gaa tta aac ct #t ttt ctt tta cat tat     1844 Asn Thr Leu Phe Ser Asn Ser Glu Leu Asn Le #u Phe Leu Leu His Tyr 405                 4 #10                 4 #15                 4 #20 ctt tcc ttg gac aat gac ttg tca aat att ct #t aaa gtg gat ttc cag     1892 Leu Ser Leu Asp Asn Asp Leu Ser Asn Ile Le #u Lys Val Asp Phe Gln                 425   #               430   #               435 aat ggt cat aac ata tgt aca ttt gca aaa tg #g tgt ata aac aac aac     1940 Asn Gly His Asn Ile Cys Thr Phe Ala Lys Tr #p Cys Ile Asn Asn Asn             440       #           445       #           450 tta gat gaa ccg tct aat tta aag cac ttt cg #t gaa atg tta gat tat     1988 Leu Asp Glu Pro Ser Asn Leu Lys His Phe Ar #g Glu Met Leu Asp Tyr         455           #       460           #       465 tat agc tct cat aat gtt aca ata agt gag ga #c gac ctg aag aac ttc     2036 Tyr Ser Ser His Asn Val Thr Ile Ser Glu As #p Asp Leu Lys Asn Phe     470               #   475               #   480 tct tta gtt ttg tgt act cat gtt gca aag gt #g aat gag aaa aca aat     2084 Ser Leu Val Leu Cys Thr His Val Ala Lys Va #l Asn Glu Lys Thr Asn 485                 4 #90                 4 #95                 5 #00 agt att ttc cgc aca tat gaa gta cat ggt tg #t gaa gtt tgt aac tca     2132 Ser Ile Phe Arg Thr Tyr Glu Val His Gly Cy #s Glu Val Cys Asn Ser                 505   #               510   #               515 ttt tgt tta cta ttt gat gag cgg tcg cct tt #t aaa att cct tat cac     2180 Phe Cys Leu Leu Phe Asp Glu Arg Ser Pro Ph #e Lys Ile Pro Tyr His             520       #           525       #           530 gaa ttg ttt tgt gca ttg cta aaa aat ccc ga #c ata att tcc tct tct     2228 Glu Leu Phe Cys Ala Leu Leu Lys Asn Pro As #p Ile Ile Ser Ser Ser         535           #       540           #       545 gtt aaa caa tca ttg ttg ctt gat ggc ttt tt #t cgg tgg agc cag cat     2276 Val Lys Gln Ser Leu Leu Leu Asp Gly Phe Ph #e Arg Trp Ser Gln His     550               #   555               #   560 tgc tca aac ttt aat aaa gaa tca atg tta ag #t tta aga gaa ttt att     2324 Cys Ser Asn Phe Asn Lys Glu Ser Met Leu Se #r Leu Arg Glu Phe Ile 565                 5 #70                 5 #75                 5 #80 atg aaa gca tta gcc agt act tca aga tgt tt #a cgt gtt gtt gct gca     2372 Met Lys Ala Leu Ala Ser Thr Ser Arg Cys Le #u Arg Val Val Ala Ala                 585   #               590   #               595 aaa gtt ttg ccc att ttc att aag gga cct aa #t aat ctt gat ata gtt     2420 Lys Val Leu Pro Ile Phe Ile Lys Gly Pro As #n Asn Leu Asp Ile Val             600       #           605       #           610 gaa ttt cac aag gaa agt aaa gcc ttg att tt #t aat acg ttg aaa ata     2468 Glu Phe His Lys Glu Ser Lys Ala Leu Ile Ph #e Asn Thr Leu Lys Ile         615           #       620           #       625 ttg gcg gtg gaa aat aca gct att tta gaa ac #g gtc att ctt tcc tgg     2516 Leu Ala Val Glu Asn Thr Ala Ile Leu Glu Th #r Val Ile Leu Ser Trp     630               #   635               #   640 atc tcc tta tct aga gtg gta gaa gaa gaa ga #a tta cat ttt gta cta     2564 Ile Ser Leu Ser Arg Val Val Glu Glu Glu Gl #u Leu His Phe Val Leu 645                 6 #50                 6 #55                 6 #60 ttg gaa gtt ata tct tct gtg ata aac agc gg #a ata ttt tat caa ggc     2612 Leu Glu Val Ile Ser Ser Val Ile Asn Ser Gl #y Ile Phe Tyr Gln Gly                 665   #               670   #               675 att ggt ctc agc gct ctg caa caa att gcc tc #g acg cgt cat ata tcc     2660 Ile Gly Leu Ser Ala Leu Gln Gln Ile Ala Se #r Thr Arg His Ile Ser             680       #           685       #           690 gtt tgg caa tta ctt tct cca tat tgg cca ac #a gtg tcc gtt gcg att     2708 Val Trp Gln Leu Leu Ser Pro Tyr Trp Pro Th #r Val Ser Val Ala Ile         695           #       700           #       705 gtc caa ggt atg ggt aaa aaa ccg aac ata gc #c agt tta ttt gct cag     2756 Val Gln Gly Met Gly Lys Lys Pro Asn Ile Al #a Ser Leu Phe Ala Gln     710               #   715               #   720 ctt atg aat att tcc gag ggc gat ttt ctt at #t cga aca cag gcg tac     2804 Leu Met Asn Ile Ser Glu Gly Asp Phe Leu Il #e Arg Thr Gln Ala Tyr 725                 7 #30                 7 #35                 7 #40 act tta cca ttc ctt gta ctt act aaa aac aa #a gcg tta ata gta cgt     2852 Thr Leu Pro Phe Leu Val Leu Thr Lys Asn Ly #s Ala Leu Ile Val Arg                 745   #               750   #               755 ata gct gaa ctt tca caa agt gat gtt gct ac #t ttg tgc ctt acc aat     2900 Ile Ala Glu Leu Ser Gln Ser Asp Val Ala Th #r Leu Cys Leu Thr Asn             760       #           765       #           770 atg cat aaa atc ctt gct tcg cta ctt act ac #g gat cat cct aat ttg     2948 Met His Lys Ile Leu Ala Ser Leu Leu Thr Th #r Asp His Pro Asn Leu         775           #       780           #       785 gaa gag agt gtg atg ctt ctt ctt tca ctg gc #c act tct gat ttt gaa     2996 Glu Glu Ser Val Met Leu Leu Leu Ser Leu Al #a Thr Ser Asp Phe Glu     790               #   795               #   800 aaa gtt gat tta acg tct ttg tta cgc tct ga #t cct att tct att act     3044 Lys Val Asp Leu Thr Ser Leu Leu Arg Ser As #p Pro Ile Ser Ile Thr 805                 8 #10                 8 #15                 8 #20 gtg gag ttg tta cag ctt tat cag aat gat gt #t cct cat gaa aaa att     3092 Val Glu Leu Leu Gln Leu Tyr Gln Asn Asp Va #l Pro His Glu Lys Ile                 825   #               830   #               835 gaa aat gct tta aga aag gta gca atg att gt #c tct caa gtg gtt aat     3140 Glu Asn Ala Leu Arg Lys Val Ala Met Ile Va #l Ser Gln Val Val Asn             840       #           845       #           850 gac gaa gac ttg agc aat aag gaa tta ctt ta #t gat ttt ttt aat aat     3188 Asp Glu Asp Leu Ser Asn Lys Glu Leu Leu Ty #r Asp Phe Phe Asn Asn         855           #       860           #       865 cac att ttg ggt atc tta gca gaa ttt tct aa #t atc ctt aac gac ctg     3236 His Ile Leu Gly Ile Leu Ala Glu Phe Ser As #n Ile Leu Asn Asp Leu     870               #   875               #   880 aaa gga aag act tca att aat gaa aag att aa #g aca att gtc ggc att     3284 Lys Gly Lys Thr Ser Ile Asn Glu Lys Ile Ly #s Thr Ile Val Gly Ile 885                 8 #90                 8 #95                 9 #00 gaa aaa atg tta tct tta tgt gga ggt gca gt #c aaa ctt gga tta cca     3332 Glu Lys Met Leu Ser Leu Cys Gly Gly Ala Va #l Lys Leu Gly Leu Pro                 905   #               910   #               915 cag ata ctt tct aat tta caa agt gct ttt ca #a aat gag cac tta agg     3380 Gln Ile Leu Ser Asn Leu Gln Ser Ala Phe Gl #n Asn Glu His Leu Arg             920       #           925       #           930 ttt tat gca atc aaa gct tgg ttc agt ttg at #a tta gca acc aag gag     3428 Phe Tyr Ala Ile Lys Ala Trp Phe Ser Leu Il #e Leu Ala Thr Lys Glu         935           #       940           #       945 ccc gag tat agt tca att gct ggt tta agt ct #t gta att tta cct cct     3476 Pro Glu Tyr Ser Ser Ile Ala Gly Leu Ser Le #u Val Ile Leu Pro Pro     950               #   955               #   960 tta ttc cct tat tta gaa cca caa gaa gca ga #g cta gta att caa ata     3524 Leu Phe Pro Tyr Leu Glu Pro Gln Glu Ala Gl #u Leu Val Ile Gln Ile 965                 9 #70                 9 #75                 9 #80 ttt gat ttt att tct tct gac aca cac aag tg #c cta caa gga tta aag     3572 Phe Asp Phe Ile Ser Ser Asp Thr His Lys Cy #s Leu Gln Gly Leu Lys                 985   #               990   #               995 tgg gct atc ccc  acc agt ctg gat tca  g #cg tgc ttt agc ctt  aag      3617 Trp Ala Ile Pro  Thr Ser Leu Asp Ser  A #la Cys Phe Ser Leu  Lys             1000      #            1005      #            1010 gct aaa gaa ata  ttc tgt tcg ctt caa  a #at gaa gat ttt tac  tct      3662 Ala Lys Glu Ile  Phe Cys Ser Leu Gln  A #sn Glu Asp Phe Tyr  Ser             1015      #            1020      #            1025 gag ctt caa agt  ata att aag tgt tta  a #ct aac gaa aat gag  cca      3707 Glu Leu Gln Ser  Ile Ile Lys Cys Leu  T #hr Asn Glu Asn Glu  Pro             1030      #            1035      #            1040 gtt tgt tat tta  ggt tta caa aaa tta  g #aa ctt ttt ttt caa  gcc      3752 Val Cys Tyr Leu  Gly Leu Gln Lys Leu  G #lu Leu Phe Phe Gln  Ala             1045      #            1050      #            1055 aag gtg gac gag  tta cat gac aca cta  a #at ttg gac ata tcc  aac      3797 Lys Val Asp Glu  Leu His Asp Thr Leu  A #sn Leu Asp Ile Ser  Asn             1060      #            1065      #            1070 gaa gtt ctg gac  caa tta cta aga tgc  c #tt tta gat tgt tgt  gta      3842 Glu Val Leu Asp  Gln Leu Leu Arg Cys  L #eu Leu Asp Cys Cys  Val             1075      #            1080      #            1085 aaa tat gct tca  aca aat atg caa ata  t #ca tat ctt gct gca  aaa      3887 Lys Tyr Ala Ser  Thr Asn Met Gln Ile  S #er Tyr Leu Ala Ala  Lys             1090      #            1095      #            1100 aat ctt ggt gaa  ttg ggt gcg ata gat  c #cc agc cgc gcc aag  gct      3932 Asn Leu Gly Glu  Leu Gly Ala Ile Asp  P #ro Ser Arg Ala Lys  Ala             1105      #            1110      #            1115 caa cat att att  aaa gaa aca gtt gtt  c #tt gat aac ttt gaa  aac      3977 Gln His Ile Ile  Lys Glu Thr Val Val  L #eu Asp Asn Phe Glu  Asn             1120      #            1125      #            1130 gga gaa gaa agt  ttg aag ttt att cta  g #at ttt atg caa tcg  cag      4022 Gly Glu Glu Ser  Leu Lys Phe Ile Leu  A #sp Phe Met Gln Ser  Gln             1135      #            1140      #            1145 tta att cca gct  ttc ctt gtt act act  g #at act aaa gca caa  ggt      4067 Leu Ile Pro Ala  Phe Leu Val Thr Thr  A #sp Thr Lys Ala Gln  Gly             1150      #            1155      #            1160 ttt ctt gcc tat  gct ctg caa gag ttt  c #ta aag ctt ggt gga  ttc      4112 Phe Leu Ala Tyr  Ala Leu Gln Glu Phe  L #eu Lys Leu Gly Gly  Phe             1165      #            1170      #            1175 aag tcc gca gtg  att aat aaa aaa aag  g #ga cta act gtg gta  aca      4157 Lys Ser Ala Val  Ile Asn Lys Lys Lys  G #ly Leu Thr Val Val  Thr             1180      #            1185      #            1190 gaa cat tgg atg  tct ttg cct gat tta  t #cc aaa cgt gtg ctt  ata      4202 Glu His Trp Met  Ser Leu Pro Asp Leu  S #er Lys Arg Val Leu  Ile             1195      #            1200      #            1205 cca ttt tta act  tcc aag tat cat tta  a #ca cca atc ccc aaa  att      4247 Pro Phe Leu Thr  Ser Lys Tyr His Leu  T #hr Pro Ile Pro Lys  Ile             1210      #            1215      #            1220 gac att cgg tac  cct att tat aaa gaa  a #at gtt act att cat  act      4292 Asp Ile Arg Tyr  Pro Ile Tyr Lys Glu  A #sn Val Thr Ile His  Thr             1225      #            1230      #            1235 tgg atg cag ttg  ttt tct ctt aaa ttg  a #tg gag tac gcc cat  tcg      4337 Trp Met Gln Leu  Phe Ser Leu Lys Leu  M #et Glu Tyr Ala His  Ser             1240      #            1245      #            1250 caa aac gct gaa  aaa ata ttt ggt att  t #gt tcg aaa gta gtg  aaa      4382 Gln Asn Ala Glu  Lys Ile Phe Gly Ile  C #ys Ser Lys Val Val  Lys             1255      #            1260      #            1265 gac caa gag gtt  aac att ccc tgt ttt  c #tt ctt ccc ttt ctt  gtt      4427 Asp Gln Glu Val  Asn Ile Pro Cys Phe  L #eu Leu Pro Phe Leu  Val             1270      #            1275      #            1280 tta aat gtt att  tta acc gag tca gaa  c #tg gaa gtt aat aaa  gtc      4472 Leu Asn Val Ile  Leu Thr Glu Ser Glu  L #eu Glu Val Asn Lys  Val             1285      #            1290      #            1295 att gaa gaa ttc  cag ctt gtt att aat  c #aa ccg gga cct gat  gga      4517 Ile Glu Glu Phe  Gln Leu Val Ile Asn  G #ln Pro Gly Pro Asp  Gly             1300      #            1305      #            1310 tta aat tcc gtg  ggg caa caa aga tac  a #cc tca ttt gta gat  gta      4562 Leu Asn Ser Val  Gly Gln Gln Arg Tyr  T #hr Ser Phe Val Asp  Val             1315      #            1320      #            1325 ttt ttt aag att  gtg gat tac ctt aac  a #aa tgg ctt cgc atg  cga      4607 Phe Phe Lys Ile  Val Asp Tyr Leu Asn  L #ys Trp Leu Arg Met  Arg             1330      #            1335      #            1340 aag aag agg aat  tgg gat aga cgt tct  g #cc att gca agg aaa  gag      4652 Lys Lys Arg Asn  Trp Asp Arg Arg Ser  A #la Ile Ala Arg Lys  Glu             1345      #            1350      #            1355 aac cgt tat atg  tcg gtg gaa gat gct  a #cc tct cga gaa tca  tcg      4697 Asn Arg Tyr Met  Ser Val Glu Asp Ala  T #hr Ser Arg Glu Ser  Ser             1360      #            1365      #            1370 atc tca aaa gtt  gag tca ttt ctt tct  c #ga ttt cct tca aaa  aca      4742 Ile Ser Lys Val  Glu Ser Phe Leu Ser  A #rg Phe Pro Ser Lys  Thr             1375      #            1380      #            1385 tta ggt att gtc  tct tta aat tgt gga  t #tt cat gct cgt gca  ttg      4787 Leu Gly Ile Val  Ser Leu Asn Cys Gly  P #he His Ala Arg Ala  Leu             1390      #            1395      #            1400 ttt tat tgg gag  caa cac ata cgt aat  g #ct aca gct cca tat  gca      4832 Phe Tyr Trp Glu  Gln His Ile Arg Asn  A #la Thr Ala Pro Tyr  Ala             1405      #            1410      #            1415 gct tta gag tcc  gat tat aga gtt ttg  c #ag gaa ata tat gct  gga      4877 Ala Leu Glu Ser  Asp Tyr Arg Val Leu  G #ln Glu Ile Tyr Ala  Gly             1420      #            1425      #            1430 att gat gat cca  gat gaa atc gaa gca  g #tg tct tta aat ttc  cat      4922 Ile Asp Asp Pro  Asp Glu Ile Glu Ala  V #al Ser Leu Asn Phe  His             1435      #            1440      #            1445 gat tac tcg ttt  gat caa caa ctc ctt  t #ta cat gaa aat tca  gga      4967 Asp Tyr Ser Phe  Asp Gln Gln Leu Leu  L #eu His Glu Asn Ser  Gly             1450      #            1455      #            1460 aca tgg gac tcg  gct ttg agt tgt tac  g #aa att att att caa  aag      5012 Thr Trp Asp Ser  Ala Leu Ser Cys Tyr  G #lu Ile Ile Ile Gln  Lys             1465      #            1470      #            1475 gat cct gaa aat  aaa aag gcg aaa atc  g #gt ttg ctt aac agc  atg      5057 Asp Pro Glu Asn  Lys Lys Ala Lys Ile  G #ly Leu Leu Asn Ser  Met             1480      #            1485      #            1490 ctg caa tcg ggg  cat tat gaa tct ctt  g #tt ttg agt tta gat  tct      5102 Leu Gln Ser Gly  His Tyr Glu Ser Leu  V #al Leu Ser Leu Asp  Ser             1495      #            1500      #            1505 ttt ata atc aat  gac aac cac gag tat  t #cg aag atg tta aat  ttg      5147 Phe Ile Ile Asn  Asp Asn His Glu Tyr  S #er Lys Met Leu Asn  Leu             1510      #            1515      #            1520 ggt att gaa gct  tca tgg cgt tcg cta  t #ct att gat tcg tta  aaa      5192 Gly Ile Glu Ala  Ser Trp Arg Ser Leu  S #er Ile Asp Ser Leu  Lys             1525      #            1530      #            1535 aag tgt ctt tca  aaa agc aac ttg gaa  t #ct ttc gaa gct aaa  ttg      5237 Lys Cys Leu Ser  Lys Ser Asn Leu Glu  S #er Phe Glu Ala Lys  Leu             1540      #            1545      #            1550 ggt agc ata ttt  tac caa tac cta cgg  a #ag gat tct ttt gct  gaa      5282 Gly Ser Ile Phe  Tyr Gln Tyr Leu Arg  L #ys Asp Ser Phe Ala  Glu             1555      #            1560      #            1565 ttg acg gag cgg  ctg caa ccc ttg tac  g #tt gat gct gct aca  gca      5327 Leu Thr Glu Arg  Leu Gln Pro Leu Tyr  V #al Asp Ala Ala Thr  Ala             1570      #            1575      #            1580 att gca aac aca  ggc gcc cat tca gcc  t #at gat tgt tat gat  att      5372 Ile Ala Asn Thr  Gly Ala His Ser Ala  T #yr Asp Cys Tyr Asp  Ile             1585      #            1590      #            1595 tta tct aag ctg  cac gca att aat gac  t #tt agt agg att gct  gaa      5417 Leu Ser Lys Leu  His Ala Ile Asn Asp  P #he Ser Arg Ile Ala  Glu             1600      #            1605      #            1610 act gac gga att  gtt tcc gac aat ctt  g #at att gtt ctt cgc  cgt      5462 Thr Asp Gly Ile  Val Ser Asp Asn Leu  A #sp Ile Val Leu Arg  Arg             1615      #            1620      #            1625 cgg ctt agc caa  gta gct ccg tac ggt  a #aa ttc aag cac caa  atc      5507 Arg Leu Ser Gln  Val Ala Pro Tyr Gly  L #ys Phe Lys His Gln  Ile             1630      #            1635      #            1640 ctg tcc act cac  tta gtt ggc tat gaa  a #aa ttt gaa aac acg  aag      5552 Leu Ser Thr His  Leu Val Gly Tyr Glu  L #ys Phe Glu Asn Thr  Lys             1645      #            1650      #            1655 aaa act gct gaa  ata tat ctc gag att  g #ca aga ata tct cga  aaa      5597 Lys Thr Ala Glu  Ile Tyr Leu Glu Ile  A #la Arg Ile Ser Arg  Lys             1660      #            1665      #            1670 aat ggt caa ttt  caa aga gcc ttc aat  g #cc atc ctc aaa gca  atg      5642 Asn Gly Gln Phe  Gln Arg Ala Phe Asn  A #la Ile Leu Lys Ala  Met             1675      #            1680      #            1685 gat tta gat aaa  ccg cta gca aca ata  g #ag cac gca caa tgg  tgg      5687 Asp Leu Asp Lys  Pro Leu Ala Thr Ile  G #lu His Ala Gln Trp  Trp             1690      #            1695      #            1700 tgg cat caa ggg  caa cat cgt aaa gct  a #tt tct gaa ttg aat  ttt      5732 Trp His Gln Gly  Gln His Arg Lys Ala  I #le Ser Glu Leu Asn  Phe             1705      #            1710      #            1715 tcg ctt aat aac  aac atg ttt gat ttg  g #tt gat gag cat gaa  gaa      5777 Ser Leu Asn Asn  Asn Met Phe Asp Leu  V #al Asp Glu His Glu  Glu             1720      #            1725      #            1730 aga cct aaa aat  cgt aaa gaa act tta  g #ga aat cca ctt aaa  gga      5822 Arg Pro Lys Asn  Arg Lys Glu Thr Leu  G #ly Asn Pro Leu Lys  Gly             1735      #            1740      #            1745 aaa gtg ttc ttg  aaa ctt aca aaa tgg  c #tc gga aaa gct ggc  caa      5867 Lys Val Phe Leu  Lys Leu Thr Lys Trp  L #eu Gly Lys Ala Gly  Gln             1750      #            1755      #            1760 ctg gga ttg aag  gat ttg gag acg tat  t #at cat aaa gcg gta  gag      5912 Leu Gly Leu Lys  Asp Leu Glu Thr Tyr  T #yr His Lys Ala Val  Glu             1765      #            1770      #            1775 att tac tca gaa  tgt gag aat acg cat  t #at tat ctt ggc cat  cat      5957 Ile Tyr Ser Glu  Cys Glu Asn Thr His  T #yr Tyr Leu Gly His  His             1780      #            1785      #            1790 cga gtt tta atg  tat gaa gaa gaa caa  a #ag ctc cca gtt aat  gaa      6002 Arg Val Leu Met  Tyr Glu Glu Glu Gln  L #ys Leu Pro Val Asn  Glu             1795      #            1800      #            1805 cag agc gaa cga  ttt tta agt ggt gag  t #ta gta act cgc ata  att      6047 Gln Ser Glu Arg  Phe Leu Ser Gly Glu  L #eu Val Thr Arg Ile  Ile             1810      #            1815      #            1820 aac gaa ttt ggt  cga tct ttg tac tat  g #gt aca aat cat ata  tat      6092 Asn Glu Phe Gly  Arg Ser Leu Tyr Tyr  G #ly Thr Asn His Ile  Tyr             1825      #            1830      #            1835 gaa agt atg cca  aaa ttg ctc aca ctg  t #gg ctt gat ttt ggg  gcc      6137 Glu Ser Met Pro  Lys Leu Leu Thr Leu  T #rp Leu Asp Phe Gly  Ala             1840      #            1845      #            1850 gaa gaa ctt cgc  tta tct aaa gat gac  g #gc gaa aag tac ttt  cgt      6182 Glu Glu Leu Arg  Leu Ser Lys Asp Asp  G #ly Glu Lys Tyr Phe  Arg             1855      #            1860      #            1865 gaa cac att atc  tct tcg aga aaa aaa  t #ct ttg gaa ctt atg  aat      6227 Glu His Ile Ile  Ser Ser Arg Lys Lys  S #er Leu Glu Leu Met  Asn             1870      #            1875      #            1880 tcg aat gtt tgt  cgc ctt tct atg aaa  a #tt cct caa tac ttt  ttt      6272 Ser Asn Val Cys  Arg Leu Ser Met Lys  I #le Pro Gln Tyr Phe  Phe             1885      #            1890      #            1895 ctg gtt gca tta  tcc caa atg ata tcc  a #ga gta tgc cat cca  aat      6317 Leu Val Ala Leu  Ser Gln Met Ile Ser  A #rg Val Cys His Pro  Asn             1900      #            1905      #            1910 aat aaa gtt tat  aaa att ttg gaa cat  a #ta att gca aac gtt  gta      6362 Asn Lys Val Tyr  Lys Ile Leu Glu His  I #le Ile Ala Asn Val  Val             1915      #            1920      #            1925 gca tct tat cct  ggg gag acg cta tgg  c #aa tta atg gca aca  ata      6407 Ala Ser Tyr Pro  Gly Glu Thr Leu Trp  G #ln Leu Met Ala Thr  Ile             1930      #            1935      #            1940 aaa tcg act tct  caa aag cgc tcg ctt  c #gt gga aaa agc att  tta      6452 Lys Ser Thr Ser  Gln Lys Arg Ser Leu  A #rg Gly Lys Ser Ile  Leu             1945      #            1950      #            1955 aat gtt tta cat  tct agg aag ctt tct  a #tg tct tcc aaa gtt  gat      6497 Asn Val Leu His  Ser Arg Lys Leu Ser  M #et Ser Ser Lys Val  Asp             1960      #            1965      #            1970 ata aaa gca ctc  agt caa tct gca att  c #tc att act gaa aag  tta      6542 Ile Lys Ala Leu  Ser Gln Ser Ala Ile  L #eu Ile Thr Glu Lys  Leu             1975      #            1980      #            1985 atc aat ttg tgc  aat aca agg att aac  a #gt aaa tct gta aaa  atg      6587 Ile Asn Leu Cys  Asn Thr Arg Ile Asn  S #er Lys Ser Val Lys  Met             1990      #            1995      #            2000 agc tta aag gat  cat ttt cgg ctt tct  t #tt gat gat ccg gta  gat      6632 Ser Leu Lys Asp  His Phe Arg Leu Ser  P #he Asp Asp Pro Val  Asp             2005      #            2010      #            2015 tta gtc att cct  gct aaa tca ttt tta  g #ac att act tta cca  gct      6677 Leu Val Ile Pro  Ala Lys Ser Phe Leu  A #sp Ile Thr Leu Pro  Ala             2020      #            2025      #            2030 aaa gat gct aac  aga gct agt cat tat  c #ca ttt cca aaa act  cag      6722 Lys Asp Ala Asn  Arg Ala Ser His Tyr  P #ro Phe Pro Lys Thr  Gln             2035      #            2040      #            2045 cct act ctg ttg  aaa ttt gag gat gag  g #tg gat ata atg aac  tct      6767 Pro Thr Leu Leu  Lys Phe Glu Asp Glu  V #al Asp Ile Met Asn  Ser             2050      #            2055      #            2060 ctt caa aaa cca  aga aaa gtg tac gtt  a #ga ggt acg gat ggc  aac      6812 Leu Gln Lys Pro  Arg Lys Val Tyr Val  A #rg Gly Thr Asp Gly  Asn             2065      #            2070      #            2075 tta tac cca ttc  ttg tgc aaa ccc aaa  g #at gat ctt cgt aag  gat      6857 Leu Tyr Pro Phe  Leu Cys Lys Pro Lys  A #sp Asp Leu Arg Lys  Asp             2080      #            2085      #            2090 gct aga ttg atg  gaa ttt aat aat ctt  a #tt tgt aaa ata ttg  agg      6902 Ala Arg Leu Met  Glu Phe Asn Asn Leu  I #le Cys Lys Ile Leu  Arg             2095      #            2100      #            2105 aaa gat caa gaa  gcg aac aga agg aac  t #tg tgt att aga act  tat      6947 Lys Asp Gln Glu  Ala Asn Arg Arg Asn  L #eu Cys Ile Arg Thr  Tyr             2110      #            2115      #            2120 gtt gtt att cct  tta aat gaa gaa tgc  g #ga ttt atc gaa tgg  gta      6992 Val Val Ile Pro  Leu Asn Glu Glu Cys  G #ly Phe Ile Glu Trp  Val             2125      #            2130      #            2135 aat cat act cgt  cca ttt aga gaa att  t #tg tta aaa agc tat  aga      7037 Asn His Thr Arg  Pro Phe Arg Glu Ile  L #eu Leu Lys Ser Tyr  Arg             2140      #            2145      #            2150 cag aaa aac att  ccc ata tca tat caa  g #aa atc aaa gtt gat  tta      7082 Gln Lys Asn Ile  Pro Ile Ser Tyr Gln  G #lu Ile Lys Val Asp  Leu             2155      #            2160      #            2165 gac ttt gca ctg  cga agt cct aac cct  g #gt gat ata ttt gaa  aag      7127 Asp Phe Ala Leu  Arg Ser Pro Asn Pro  G #ly Asp Ile Phe Glu  Lys             2170      #            2175      #            2180 aaa atc tta ccg  aaa ttt cct cca gtt  t #tt tat gag tgg ttt  gtt      7172 Lys Ile Leu Pro  Lys Phe Pro Pro Val  P #he Tyr Glu Trp Phe  Val             2185      #            2190      #            2195 gaa tct ttc cca  gaa cca aat aat tgg  g #tt act agt aga caa  aac      7217 Glu Ser Phe Pro  Glu Pro Asn Asn Trp  V #al Thr Ser Arg Gln  Asn             2200      #            2205      #            2210 tat tgc cga act  tta gca gta atg tca  a #ta gtt ggc tac gtt  ttg      7262 Tyr Cys Arg Thr  Leu Ala Val Met Ser  I #le Val Gly Tyr Val  Leu             2215      #            2220      #            2225 ggt ttg gga gat  cgc cat ggc gaa aac  a #ta ttg ttt gat gaa  ttt      7307 Gly Leu Gly Asp  Arg His Gly Glu Asn  I #le Leu Phe Asp Glu  Phe             2230      #            2235      #            2240 aca ggt gaa gct  atc cat gtc gat ttc  a #ac tgt ctt ttt gat  aaa      7352 Thr Gly Glu Ala  Ile His Val Asp Phe  A #sn Cys Leu Phe Asp  Lys             2245      #            2250      #            2255 ggt ctt act ttt  gaa aaa cct gaa aag  g #tg ccg ttc aga tta  act      7397 Gly Leu Thr Phe  Glu Lys Pro Glu Lys  V #al Pro Phe Arg Leu  Thr             2260      #            2265      #            2270 cat aat atg gta  gat gca atg ggt ccg  a #ca ggt tat gaa ggg  ggt      7442 His Asn Met Val  Asp Ala Met Gly Pro  T #hr Gly Tyr Glu Gly  Gly             2275      #            2280      #            2285 ttc agg aaa gct  agc gaa ata acg atg  c #gg ctt ctt cgc tca  aac      7487 Phe Arg Lys Ala  Ser Glu Ile Thr Met  A #rg Leu Leu Arg Ser  Asn             2290      #            2295      #            2300 caa gat aca ttg  atg agc gta cta gag  t #ct ttc cta cat gat  cct      7532 Gln Asp Thr Leu  Met Ser Val Leu Glu  S #er Phe Leu His Asp  Pro             2305      #            2310      #            2315 tta gtc gag tgg  aat aga aag aag tcg  t #ca agc aag tac ccg  aat      7577 Leu Val Glu Trp  Asn Arg Lys Lys Ser  S #er Ser Lys Tyr Pro  Asn             2320      #            2325      #            2330 aat gaa gca aat  gaa gtt ttg gat ata  a #tt cgc aaa aaa ttt  caa      7622 Asn Glu Ala Asn  Glu Val Leu Asp Ile  I #le Arg Lys Lys Phe  Gln             2335      #            2340      #            2345 ggc ttt atg cca  ggg gag acg ata cct  t #ta tct att gaa ggg  caa      7667 Gly Phe Met Pro  Gly Glu Thr Ile Pro  L #eu Ser Ile Glu Gly  Gln             2350      #            2355      #            2360 att caa gaa ttg  atc aaa tct gct gtc  a #ac cca aaa aac ctg  gta      7712 Ile Gln Glu Leu  Ile Lys Ser Ala Val  A #sn Pro Lys Asn Leu  Val             2365      #            2370      #            2375 gaa atg tac att  ggt tgg gct gct tat  t #tc tagcatttta ctaacaaaaa     7762 Glu Met Tyr Ile  Gly Trp Ala Ala Tyr  P #he             2380      #            2385 tttcaatgaa caagctaccc attattaaac ttatgatttg aatcgaagat at #tttattta   7822 ttaatccgat gaagaattct cgctgagttg ttcaatttct tgtaattttc ct #tccatttc   7882 taaatcgtcg attcgcttaa atagggcact ggctttttgt gcatttttct ct #cgtaaagc   7942 agcttctgat tgaaaaaaag ctatatctgt ttctgagtca tcatccgaat ca #acaatata   8002 ttttgcagat cgacctgcag             #                   #                 802 #2 <210> SEQ ID NO 4 <211> LENGTH: 2386 <212> TYPE: PRT <213> ORGANISM: Homo sapiens <400> SEQUENCE: 4 Met Ser Gln His Ala Lys Arg Lys Ala Gly Se #r Leu Asp Leu Ser Pro 1               5    #                10   #                15 Arg Gly Leu Asp Asp Arg Gln Ala Phe Gly Gl #n Leu Leu Lys Glu Val             20       #            25       #            30 Leu Ala Leu Asp Lys Glu His Glu Leu Gly Ar #g Ser Asn Ser Leu Pro         35           #        40           #        45 Ser Met Thr Ser Glu Leu Val Glu Val Leu Il #e Glu Val Gly Leu Leu     50               #    55               #    60 Ala Phe Lys His Asp Asp Ser Lys Ser Glu Ph #e Ile Ser Pro Lys Met 65                   #70                   #75                   #80 Leu Lys Glu Ala His Leu Ser Leu Gln Ala Le #u Met Leu Ile Leu Lys                 85   #                90   #                95 Arg Ser Pro Thr Val Leu Arg Glu Ile Lys Se #r Ser Val Thr Leu Leu             100       #           105       #           110 Asp Trp Ile Leu Pro Arg Thr Ile Ser Leu Ph #e Ala Asp Ile Arg Phe         115           #       120           #       125 Ile Lys Leu Phe Asp Ser Leu Lys Glu Phe Hi #s Lys Leu Ile Tyr Gln     130               #   135               #   140 Leu Ile Ser Glu Lys Ser Phe Leu Trp Asp Le #u Tyr Ala Ser Phe Met 145                 1 #50                 1 #55                 1 #60 Arg Tyr Trp Lys Tyr Tyr Ile Thr Asn Val Se #r Ser Ile Val Leu Gln                 165   #               170   #               175 Ile Thr Asn Ala Thr Phe Pro Tyr Lys Met Pr #o Ser Pro Asn Ser Gln             180       #           185       #           190 Pro Leu Gln Ser Ile Ser Pro Asn Tyr Pro Th #r His Arg Glu Asp Lys         195           #       200           #       205 Phe Asp Leu Leu Ile Ile Asn Ile Glu Glu Al #a Cys Thr Phe Phe Phe     210               #   215               #   220 Glu Ser Ala His Phe Phe Ala Gln Cys Ser Ty #r Leu Lys Lys Ser Asn 225                 2 #30                 2 #35                 2 #40 Phe Pro Ser Pro Pro Leu Phe Thr Ala Trp Th #r Trp Ile Lys Pro Cys                 245   #               250   #               255 Phe Phe Asn Phe Val Ile Leu Leu Lys Arg Il #e Ser Ile Gly Asp Ser             260       #           265       #           270 Gln Leu Phe Leu His Leu His Ser Arg Ile Va #l Gln Thr Leu Cys Cys         275           #       280           #       285 Phe Ser Leu Asn Phe Ile Tyr His Gly Leu Pr #o Ile Cys Glu Lys Ser     290               #   295               #   300 Lys His Ile Leu Met Ser Ser Ile Asn Leu Th #r Leu Gly Ser Leu Lys 305                 3 #10                 3 #15                 3 #20 Lys Thr Tyr Thr Val Ala Asn Thr Ala Ile Se #r Leu Phe Phe Leu Ser                 325   #               330   #               335 Leu Phe Val Leu Pro Lys Thr Val Ala Gly Le #u Phe Tyr Pro Phe Gly             340       #           345       #           350 Val Ser Leu Leu Ser Asp Phe Lys Val Leu Gl #u Gln Leu Glu Pro Asp         355           #       360           #       365 Ser Asp Leu Lys Lys Ala Ile Ile Leu Phe Ly #s Cys Arg Tyr Gln Ser     370               #   375               #   380 Ser Glu Ile Asp Gln Thr Thr Leu Arg Ala Ph #e Gly Glu Ile Cys Thr 385                 3 #90                 3 #95                 4 #00 Gly Lys Leu Glu Asn Thr Leu Phe Ser Asn Se #r Glu Leu Asn Leu Phe                 405   #               410   #               415 Leu Leu His Tyr Leu Ser Leu Asp Asn Asp Le #u Ser Asn Ile Leu Lys             420       #           425       #           430 Val Asp Phe Gln Asn Gly His Asn Ile Cys Th #r Phe Ala Lys Trp Cys         435           #       440           #       445 Ile Asn Asn Asn Leu Asp Glu Pro Ser Asn Le #u Lys His Phe Arg Glu     450               #   455               #   460 Met Leu Asp Tyr Tyr Ser Ser His Asn Val Th #r Ile Ser Glu Asp Asp 465                 4 #70                 4 #75                 4 #80 Leu Lys Asn Phe Ser Leu Val Leu Cys Thr Hi #s Val Ala Lys Val Asn                 485   #               490   #               495 Glu Lys Thr Asn Ser Ile Phe Arg Thr Tyr Gl #u Val His Gly Cys Glu             500       #           505       #           510 Val Cys Asn Ser Phe Cys Leu Leu Phe Asp Gl #u Arg Ser Pro Phe Lys         515           #       520           #       525 Ile Pro Tyr His Glu Leu Phe Cys Ala Leu Le #u Lys Asn Pro Asp Ile     530               #   535               #   540 Ile Ser Ser Ser Val Lys Gln Ser Leu Leu Le #u Asp Gly Phe Phe Arg 545                 5 #50                 5 #55                 5 #60 Trp Ser Gln His Cys Ser Asn Phe Asn Lys Gl #u Ser Met Leu Ser Leu                 565   #               570   #               575 Arg Glu Phe Ile Met Lys Ala Leu Ala Ser Th #r Ser Arg Cys Leu Arg             580       #           585       #           590 Val Val Ala Ala Lys Val Leu Pro Ile Phe Il #e Lys Gly Pro Asn Asn         595           #       600           #       605 Leu Asp Ile Val Glu Phe His Lys Glu Ser Ly #s Ala Leu Ile Phe Asn     610               #   615               #   620 Thr Leu Lys Ile Leu Ala Val Glu Asn Thr Al #a Ile Leu Glu Thr Val 625                 6 #30                 6 #35                 6 #40 Ile Leu Ser Trp Ile Ser Leu Ser Arg Val Va #l Glu Glu Glu Glu Leu                 645   #               650   #               655 His Phe Val Leu Leu Glu Val Ile Ser Ser Va #l Ile Asn Ser Gly Ile             660       #           665       #           670 Phe Tyr Gln Gly Ile Gly Leu Ser Ala Leu Gl #n Gln Ile Ala Ser Thr         675           #       680           #       685 Arg His Ile Ser Val Trp Gln Leu Leu Ser Pr #o Tyr Trp Pro Thr Val     690               #   695               #   700 Ser Val Ala Ile Val Gln Gly Met Gly Lys Ly #s Pro Asn Ile Ala Ser 705                 7 #10                 7 #15                 7 #20 Leu Phe Ala Gln Leu Met Asn Ile Ser Glu Gl #y Asp Phe Leu Ile Arg                 725   #               730   #               735 Thr Gln Ala Tyr Thr Leu Pro Phe Leu Val Le #u Thr Lys Asn Lys Ala             740       #           745       #           750 Leu Ile Val Arg Ile Ala Glu Leu Ser Gln Se #r Asp Val Ala Thr Leu         755           #       760           #       765 Cys Leu Thr Asn Met His Lys Ile Leu Ala Se #r Leu Leu Thr Thr Asp     770               #   775               #   780 His Pro Asn Leu Glu Glu Ser Val Met Leu Le #u Leu Ser Leu Ala Thr 785                 7 #90                 7 #95                 8 #00 Ser Asp Phe Glu Lys Val Asp Leu Thr Ser Le #u Leu Arg Ser Asp Pro                 805   #               810   #               815 Ile Ser Ile Thr Val Glu Leu Leu Gln Leu Ty #r Gln Asn Asp Val Pro             820       #           825       #           830 His Glu Lys Ile Glu Asn Ala Leu Arg Lys Va #l Ala Met Ile Val Ser         835           #       840           #       845 Gln Val Val Asn Asp Glu Asp Leu Ser Asn Ly #s Glu Leu Leu Tyr Asp     850               #   855               #   860 Phe Phe Asn Asn His Ile Leu Gly Ile Leu Al #a Glu Phe Ser Asn Ile 865                 8 #70                 8 #75                 8 #80 Leu Asn Asp Leu Lys Gly Lys Thr Ser Ile As #n Glu Lys Ile Lys Thr                 885   #               890   #               895 Ile Val Gly Ile Glu Lys Met Leu Ser Leu Cy #s Gly Gly Ala Val Lys             900       #           905       #           910 Leu Gly Leu Pro Gln Ile Leu Ser Asn Leu Gl #n Ser Ala Phe Gln Asn         915           #       920           #       925 Glu His Leu Arg Phe Tyr Ala Ile Lys Ala Tr #p Phe Ser Leu Ile Leu     930               #   935               #   940 Ala Thr Lys Glu Pro Glu Tyr Ser Ser Ile Al #a Gly Leu Ser Leu Val 945                 9 #50                 9 #55                 9 #60 Ile Leu Pro Pro Leu Phe Pro Tyr Leu Glu Pr #o Gln Glu Ala Glu Leu                 965   #               970   #               975 Val Ile Gln Ile Phe Asp Phe Ile Ser Ser As #p Thr His Lys Cys Leu             980       #           985       #           990 Gln Gly Leu Lys Trp Ala Ile Pro  Thr Ser  #Leu Asp Ser  Ala Cys Phe         995           #       1000           #       1005 Ser Leu  Lys Ala Lys Glu Ile  Phe Cys S #er Leu Gln  Asn Glu Asp     1010              #    1015              #    1020 Phe Tyr  Ser Glu Leu Gln Ser  Ile Ile L #ys Cys Leu  Thr Asn Glu     1025              #    1030              #    1035 Asn Glu  Pro Val Cys Tyr Leu  Gly Leu G #ln Lys Leu  Glu Leu Phe     1040              #    1045              #    1050 Phe Gln  Ala Lys Val Asp Glu  Leu His A #sp Thr Leu  Asn Leu Asp     1055              #    1060              #    1065 Ile Ser  Asn Glu Val Leu Asp  Gln Leu L #eu Arg Cys  Leu Leu Asp     1070              #    1075              #    1080 Cys Cys  Val Lys Tyr Ala Ser  Thr Asn M #et Gln Ile  Ser Tyr Leu     1085              #    1090              #    1095 Ala Ala  Lys Asn Leu Gly Glu  Leu Gly A #la Ile Asp  Pro Ser Arg     1100              #    1105              #    1110 Ala Lys  Ala Gln His Ile Ile  Lys Glu T #hr Val Val  Leu Asp Asn     1115              #    1120              #    1125 Phe Glu  Asn Gly Glu Glu Ser  Leu Lys P #he Ile Leu  Asp Phe Met     1130              #    1135              #    1140 Gln Ser  Gln Leu Ile Pro Ala  Phe Leu V #al Thr Thr  Asp Thr Lys     1145              #    1150              #    1155 Ala Gln  Gly Phe Leu Ala Tyr  Ala Leu G #ln Glu Phe  Leu Lys Leu     1160              #    1165              #    1170 Gly Gly  Phe Lys Ser Ala Val  Ile Asn L #ys Lys Lys  Gly Leu Thr     1175              #    1180              #    1185 Val Val  Thr Glu His Trp Met  Ser Leu P #ro Asp Leu  Ser Lys Arg     1190              #    1195              #    1200 Val Leu  Ile Pro Phe Leu Thr  Ser Lys T #yr His Leu  Thr Pro Ile     1205              #    1210              #    1215 Pro Lys  Ile Asp Ile Arg Tyr  Pro Ile T #yr Lys Glu  Asn Val Thr     1220              #    1225              #    1230 Ile His  Thr Trp Met Gln Leu  Phe Ser L #eu Lys Leu  Met Glu Tyr     1235              #    1240              #    1245 Ala His  Ser Gln Asn Ala Glu  Lys Ile P #he Gly Ile  Cys Ser Lys     1250              #    1255              #    1260 Val Val  Lys Asp Gln Glu Val  Asn Ile P #ro Cys Phe  Leu Leu Pro     1265              #    1270              #    1275 Phe Leu  Val Leu Asn Val Ile  Leu Thr G #lu Ser Glu  Leu Glu Val     1280              #    1285              #    1290 Asn Lys  Val Ile Glu Glu Phe  Gln Leu V #al Ile Asn  Gln Pro Gly     1295              #    1300              #    1305 Pro Asp  Gly Leu Asn Ser Val  Gly Gln G #ln Arg Tyr  Thr Ser Phe     1310              #    1315              #    1320 Val Asp  Val Phe Phe Lys Ile  Val Asp T #yr Leu Asn  Lys Trp Leu     1325              #    1330              #    1335 Arg Met  Arg Lys Lys Arg Asn  Trp Asp A #rg Arg Ser  Ala Ile Ala     1340              #    1345              #    1350 Arg Lys  Glu Asn Arg Tyr Met  Ser Val G #lu Asp Ala  Thr Ser Arg     1355              #    1360              #    1365 Glu Ser  Ser Ile Ser Lys Val  Glu Ser P #he Leu Ser  Arg Phe Pro     1370              #    1375              #    1380 Ser Lys  Thr Leu Gly Ile Val  Ser Leu A #sn Cys Gly  Phe His Ala     1385              #    1390              #    1395 Arg Ala  Leu Phe Tyr Trp Glu  Gln His I #le Arg Asn  Ala Thr Ala     1400              #    1405              #    1410 Pro Tyr  Ala Ala Leu Glu Ser  Asp Tyr A #rg Val Leu  Gln Glu Ile     1415              #    1420              #    1425 Tyr Ala  Gly Ile Asp Asp Pro  Asp Glu I #le Glu Ala  Val Ser Leu     1430              #    1435              #    1440 Asn Phe  His Asp Tyr Ser Phe  Asp Gln G #ln Leu Leu  Leu His Glu     1445              #    1450              #    1455 Asn Ser  Gly Thr Trp Asp Ser  Ala Leu S #er Cys Tyr  Glu Ile Ile     1460              #    1465              #    1470 Ile Gln  Lys Asp Pro Glu Asn  Lys Lys A #la Lys Ile  Gly Leu Leu     1475              #    1480              #    1485 Asn Ser  Met Leu Gln Ser Gly  His Tyr G #lu Ser Leu  Val Leu Ser     1490              #    1495              #    1500 Leu Asp  Ser Phe Ile Ile Asn  Asp Asn H #is Glu Tyr  Ser Lys Met     1505              #    1510              #    1515 Leu Asn  Leu Gly Ile Glu Ala  Ser Trp A #rg Ser Leu  Ser Ile Asp     1520              #    1525              #    1530 Ser Leu  Lys Lys Cys Leu Ser  Lys Ser A #sn Leu Glu  Ser Phe Glu     1535              #    1540              #    1545 Ala Lys  Leu Gly Ser Ile Phe  Tyr Gln T #yr Leu Arg  Lys Asp Ser     1550              #    1555              #    1560 Phe Ala  Glu Leu Thr Glu Arg  Leu Gln P #ro Leu Tyr  Val Asp Ala     1565              #    1570              #    1575 Ala Thr  Ala Ile Ala Asn Thr  Gly Ala H #is Ser Ala  Tyr Asp Cys     1580              #    1585              #    1590 Tyr Asp  Ile Leu Ser Lys Leu  His Ala I #le Asn Asp  Phe Ser Arg     1595              #    1600              #    1605 Ile Ala  Glu Thr Asp Gly Ile  Val Ser A #sp Asn Leu  Asp Ile Val     1610              #    1615              #    1620 Leu Arg  Arg Arg Leu Ser Gln  Val Ala P #ro Tyr Gly  Lys Phe Lys     1625              #    1630              #    1635 His Gln  Ile Leu Ser Thr His  Leu Val G #ly Tyr Glu  Lys Phe Glu     1640              #    1645              #    1650 Asn Thr  Lys Lys Thr Ala Glu  Ile Tyr L #eu Glu Ile  Ala Arg Ile     1655              #    1660              #    1665 Ser Arg  Lys Asn Gly Gln Phe  Gln Arg A #la Phe Asn  Ala Ile Leu     1670              #    1675              #    1680 Lys Ala  Met Asp Leu Asp Lys  Pro Leu A #la Thr Ile  Glu His Ala     1685              #    1690              #    1695 Gln Trp  Trp Trp His Gln Gly  Gln His A #rg Lys Ala  Ile Ser Glu     1700              #    1705              #    1710 Leu Asn  Phe Ser Leu Asn Asn  Asn Met P #he Asp Leu  Val Asp Glu     1715              #    1720              #    1725 His Glu  Glu Arg Pro Lys Asn  Arg Lys G #lu Thr Leu  Gly Asn Pro     1730              #    1735              #    1740 Leu Lys  Gly Lys Val Phe Leu  Lys Leu T #hr Lys Trp  Leu Gly Lys     1745              #    1750              #    1755 Ala Gly  Gln Leu Gly Leu Lys  Asp Leu G #lu Thr Tyr  Tyr His Lys     1760              #    1765              #    1770 Ala Val  Glu Ile Tyr Ser Glu  Cys Glu A #sn Thr His  Tyr Tyr Leu     1775              #    1780              #    1785 Gly His  His Arg Val Leu Met  Tyr Glu G #lu Glu Gln  Lys Leu Pro     1790              #    1795              #    1800 Val Asn  Glu Gln Ser Glu Arg  Phe Leu S #er Gly Glu  Leu Val Thr     1805              #    1810              #    1815 Arg Ile  Ile Asn Glu Phe Gly  Arg Ser L #eu Tyr Tyr  Gly Thr Asn     1820              #    1825              #    1830 His Ile  Tyr Glu Ser Met Pro  Lys Leu L #eu Thr Leu  Trp Leu Asp     1835              #    1840              #    1845 Phe Gly  Ala Glu Glu Leu Arg  Leu Ser L #ys Asp Asp  Gly Glu Lys     1850              #    1855              #    1860 Tyr Phe  Arg Glu His Ile Ile  Ser Ser A #rg Lys Lys  Ser Leu Glu     1865              #    1870              #    1875 Leu Met  Asn Ser Asn Val Cys  Arg Leu S #er Met Lys  Ile Pro Gln     1880              #    1885              #    1890 Tyr Phe  Phe Leu Val Ala Leu  Ser Gln M #et Ile Ser  Arg Val Cys     1895              #    1900              #    1905 His Pro  Asn Asn Lys Val Tyr  Lys Ile L #eu Glu His  Ile Ile Ala     1910              #    1915              #    1920 Asn Val  Val Ala Ser Tyr Pro  Gly Glu T #hr Leu Trp  Gln Leu Met     1925              #    1930              #    1935 Ala Thr  Ile Lys Ser Thr Ser  Gln Lys A #rg Ser Leu  Arg Gly Lys     1940              #    1945              #    1950 Ser Ile  Leu Asn Val Leu His  Ser Arg L #ys Leu Ser  Met Ser Ser     1955              #    1960              #    1965 Lys Val  Asp Ile Lys Ala Leu  Ser Gln S #er Ala Ile  Leu Ile Thr     1970              #    1975              #    1980 Glu Lys  Leu Ile Asn Leu Cys  Asn Thr A #rg Ile Asn  Ser Lys Ser     1985              #    1990              #    1995 Val Lys  Met Ser Leu Lys Asp  His Phe A #rg Leu Ser  Phe Asp Asp     2000              #    2005              #    2010 Pro Val  Asp Leu Val Ile Pro  Ala Lys S #er Phe Leu  Asp Ile Thr     2015              #    2020              #    2025 Leu Pro  Ala Lys Asp Ala Asn  Arg Ala S #er His Tyr  Pro Phe Pro     2030              #    2035              #    2040 Lys Thr  Gln Pro Thr Leu Leu  Lys Phe G #lu Asp Glu  Val Asp Ile     2045              #    2050              #    2055 Met Asn  Ser Leu Gln Lys Pro  Arg Lys V #al Tyr Val  Arg Gly Thr     2060              #    2065              #    2070 Asp Gly  Asn Leu Tyr Pro Phe  Leu Cys L #ys Pro Lys  Asp Asp Leu     2075              #    2080              #    2085 Arg Lys  Asp Ala Arg Leu Met  Glu Phe A #sn Asn Leu  Ile Cys Lys     2090              #    2095              #    2100 Ile Leu  Arg Lys Asp Gln Glu  Ala Asn A #rg Arg Asn  Leu Cys Ile     2105              #    2110              #    2115 Arg Thr  Tyr Val Val Ile Pro  Leu Asn G #lu Glu Cys  Gly Phe Ile     2120              #    2125              #    2130 Glu Trp  Val Asn His Thr Arg  Pro Phe A #rg Glu Ile  Leu Leu Lys     2135              #    2140              #    2145 Ser Tyr  Arg Gln Lys Asn Ile  Pro Ile S #er Tyr Gln  Glu Ile Lys     2150              #    2155              #    2160 Val Asp  Leu Asp Phe Ala Leu  Arg Ser P #ro Asn Pro  Gly Asp Ile     2165              #    2170              #    2175 Phe Glu  Lys Lys Ile Leu Pro  Lys Phe P #ro Pro Val  Phe Tyr Glu     2180              #    2185              #    2190 Trp Phe  Val Glu Ser Phe Pro  Glu Pro A #sn Asn Trp  Val Thr Ser     2195              #    2200              #    2205 Arg Gln  Asn Tyr Cys Arg Thr  Leu Ala V #al Met Ser  Ile Val Gly     2210              #    2215              #    2220 Tyr Val  Leu Gly Leu Gly Asp  Arg His G #ly Glu Asn  Ile Leu Phe     2225              #    2230              #    2235 Asp Glu  Phe Thr Gly Glu Ala  Ile His V #al Asp Phe  Asn Cys Leu     2240              #    2245              #    2250 Phe Asp  Lys Gly Leu Thr Phe  Glu Lys P #ro Glu Lys  Val Pro Phe     2255              #    2260              #    2265 Arg Leu  Thr His Asn Met Val  Asp Ala M #et Gly Pro  Thr Gly Tyr     2270              #    2275              #    2280 Glu Gly  Gly Phe Arg Lys Ala  Ser Glu I #le Thr Met  Arg Leu Leu     2285              #    2290              #    2295 Arg Ser  Asn Gln Asp Thr Leu  Met Ser V #al Leu Glu  Ser Phe Leu     2300              #    2305              #    2310 His Asp  Pro Leu Val Glu Trp  Asn Arg L #ys Lys Ser  Ser Ser Lys     2315              #    2320              #    2325 Tyr Pro  Asn Asn Glu Ala Asn  Glu Val L #eu Asp Ile  Ile Arg Lys     2330              #    2335              #    2340 Lys Phe  Gln Gly Phe Met Pro  Gly Glu T #hr Ile Pro  Leu Ser Ile     2345              #    2350              #    2355 Glu Gly  Gln Ile Gln Glu Leu  Ile Lys S #er Ala Val  Asn Pro Lys     2360              #    2365              #    2370 Asn Leu  Val Glu Met Tyr Ile  Gly Trp A #la Ala Tyr  Phe     2375              #    2380              #    2385 <210> SEQ ID NO 5 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 5 gttttcgcca tggcgcgctc ccaaacccaa          #                   #           30 <210> SEQ ID NO 6 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 6 ttcatcaaac aatatctttt cgccatggcg          #                   #           30 <210> SEQ ID NO 7 <211> LENGTH: 30 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 7 caaaaagaca gttgaattcg acatggatag          #                   #           30 <210> SEQ ID NO 8 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 8 gacgcagaat tcaccagtca aagaatcaaa gag        #                   #         33 <210> SEQ ID NO 9 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 9 tggatgatga cagctgtgtc             #                   #                   # 20 <210> SEQ ID NO 10 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 10 tgtagtcgct gctcaatgtc             #                   #                   # 20 <210> SEQ ID NO 11 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Primer <400> SEQUENCE: 11 tggtttctga gaacattccc tga            #                   #                23 <210> SEQ ID NO 12 <211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: Rad3/Esrlp <221> NAME/KEY: MISC_FEATURE <222> LOCATION: (5)..(5) <223> OTHER INFORMATION: Xaa = Isoleucine or  #Valine <221> NAME/KEY: MISC_FEATURE <222> LOCATION: (6)..(6) <223> OTHER INFORMATION: Xaa = Leucine or Ph #enylalanine <221> NAME/KEY: MISC_FEATURE <222> LOCATION: (8)..(8) <223> OTHER INFORMATION: Xaa = Glutamine or  #Glutamic Acid <400> SEQUENCE: 12 Lys Phe Pro Pro Xaa Xaa Tyr Xaa Trp Phe 1               5    #                10 <210> SEQ ID NO 13 <211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: oligonucleutide oDH18 <400> SEQUENCE: 13 Leu Gly Leu Gly Asp Arg His 1               5 <210> SEQ ID NO 14 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Artificial sequence <220> FEATURE: <223> OTHER INFORMATION: oligonucleotide oDH-16 <221> NAME/KEY: MISC_FEATURE <222> LOCATION: (5)..(5) <223> OTHER INFORMATION: Xaa = Aspartic Acid  #or Asparagine <400> SEQUENCE: 14 His Val Asp Phe Xaa Cys 1               5 

What is claimed is:
 1. A purified and isolated polynucleotide encoding an ATR polypeptide that has lipid kinase activity, said polynucleotide selected from the group consisting of: (a) a polynucleotide encoding the amino acid sequence set out in SEQ ID NO: 2, (b) a polynucleotide encoding the amino acid sequence set out in SEQ ID NO: 4, and (c) a polynucleotide which hybridizes to the complement of the polynucleotide of (a) or (b) under conditions including a final wash at 55° C.
 2. A purified and isolated polynucleotide encoding an ATR polypeptide that has lipid kinase activity, said polynucleotide selected from the group consisting of: a) the polynucleotide set out in SEQ ID NO: 1; b) the polynucleotide set out in SEQ ID NO: 3; and c) a polynucleotide that hybridizes to the complement of the polynucleotide of (a) or (b) under conditions including a final wash at 55° C.
 3. An expression vector comprising the polynucleotide according to claim 1 or
 2. 4. An isolated host cell transformed or transfected with a polynucleotide according to claim 1 or
 2. 5. A method for producing an ATR polypeptide comprising the step of growing the host cell of claim 4 under conditions in which expression of the ATR polypeptide occurs.
 6. A method for detecting the presence of a polynucleotide according to claim 1 or 2 in a human or animal body sample comprising the steps of (a) contacting a human or animal body sample with a probe comprising a polynucleotide according to claim 1 or 2 under conditions which permit hybridization; and (b) detecting in the sample any duplex formed between the probe and a nucleic acid. 